Publications

porcine willebrand factor: a population of multimers.	purified porcine willebrand factor was analyzed by agarose-sodium dodso4 electrophoresis. multiple forms of the protein were found in a series of increasing molecular weights. a molecular mass calibration curve was constructed with fibrinogen (3.4 x 10(5) daltons), igm (1 x 10(6) daltons), and glutaraldehyde-crosslinked igm polymers (2, 3, and 4 x 10(6) daltons). as measured by this procedure, the apparent molecular weight of willebrand factor polymers ranged from 1.1 x 10(6) to 2.1 x 10(7). eac ...	1978	413873
separation of oligo-rna by reverse-phase hplc.	a rapid and highly reproducible chromatographic technique has been developed for analysis and purification of complex mixtures of oligoribonucleotides. the method utilizes a column of microparticulate porous silica beads fully derivatized with octadecylsilyl groups. the column is eluted with gradients in acetonitrile/water/ammonium acetate pumped at pressures of 1500-300 psi. most separations are completed in 5-15 min. with usually better than 1% reproducibility of absolute retention times and a ...	1979	503849
polyribosome size analysis. measurement of number-average polyribosome sizes.	the analysis of translational efficiencies of specific mrnas requires a determination of the polyribosome size. the appropriate value to use in such calculations is the number-average size. a method is described for accurately measuring the number-average size of total and of specific protein synthesizing polyribosomes using isokinetic sucrose density gradients and 125i-labeled antibodies. by this method, we demonstrated that albumin synthesizing polyribosomes from a serum albumin secreting mous ...	1979	534642
nuclear magnetic resonance investigation of the base-pairing structure of escherichia coli trnatyr monomer and dimer conformations.	the structures of the escherichia coli tyrosine trna monomer and dimer have been investigated by high-resolution nuclear magnetic resonance (nmr). at 23 degrees c the monomer contains 26 +/- 2 base pairs and the low-field nmr spectrum (11.7-15 ppm) can be accounted for in terms of the cloverleaf structure (23 base pairs) and three additional resonances that are assigned to tertiary structure base pairs. assignments suggested for the various resonances are consistent with thermal denaturation stu ...	1976	782517
pmr studies of the self-association of dna intercalating ellipticine derivatives in aqueous solution.	in aqueous solution dna intercalating ellipticine derivatives aggregate in n-mers. the self-association constants k are higher than those of 2-methoxy-6-chloro-9-[3-dimethylaminopropyl-amino]-acridine and ethidium bromide. they are of the same order as that of actinomycin d but inferior to that of acridine orange. the increase of the 9-hydroxy-ellipticine constant by addition of sodium chloride shows the importance of anion participation in the mechanism of stacking in accordance with the high e ...	1976	949529
clays as possible catalysts for peptide formation in the prebiotic era.	from the point of view of prebiotic synthesis, clays might have performed functions of concentration, catalysis, and protection of molecules. the degree of polymerization obtained when amino acid adenylates are added to montmorillonite suspensions in water, are much iigher than those obtained by polymerization in the absence of such a clay. in addition, they are of a discrete spectrum, usually multiplies of 6 or 7, and reach values of up to 40 mers. in the absence of clay a continuous spectrum o ...	1976	1023136
analysis of the two steps in polypeptide chain initiation inhibited by pactamycin.	earlier work has shown that the inhibition by pactamycin (pm) of polypeptide chain initiation in reticulocyte extracts is associated with (1) a defect in the joining of the 60s subunit to the smaller initiation complex to form an 80s complex ("joining reaction") (kappen, l. s., suzuki, h., and goldberg, i. h. (1973), proc. natl. acad. sci. u.s.a. 70, 22) and (2) a block after the synthesis of the initial dipeptide (kappen, l. s., and goldberg, i. h. (1973), biochem. biophys. res. commun. 54, 108 ...	1976	1247535
ubiquitin genes are differentially regulated in protoplast-derived cultures of nicotiana sylvestris and in response to various stresses.	four ubiquitin mrna size classes were found to be differentially regulated in mesophyll protoplast-derived cultures of nicotiana sylvestris. three mrna families of 1.9, 1.6 and 1.35 kb were expressed as soon as protoplasts were isolated. the 1.9 and 1.6 kb size classes were transiently expressed during the first hours of culture, whereas the level of expression of the 1.35 kb size class was maintained as long as cells kept dividing. a 0.7 kb mrna size class started to be expressed just before th ...	1992	1281439
use of the multipin peptide synthesis technique for the generation of antipeptide sera.	the multipin peptide synthesis technique has been used to map antigenic sites of proteins (1,2). antibodies raised to the whole protein are screened on pin-synthesized overlapping octapeptides homologous with the protein of interest, and the peptides that bind antibodies clearly identify the epitopes. what is described in this study is a method using pin-synthesized peptides to generate specific antibodies to many peptides. cleavable linkers have been developed (3) that, used together with the m ...	2003	1285329
inhibition of expression of a mouse alpha-globin gene by plasmids that include antisense oligonucleotides.	plasmid-borne dnas, corresponding to 68-base oligodeoxynucleotides, synthesized in the antisense or sense configuration and based on the nucleotide sequences of various regions of the mouse alpha-globin mrna, were introduced with the gene for xanthine-guanine phosphoribosyl transferase from e. coli (ecogpt) into mouse erythroleukemia (mel) cells by protoplast fusion. specific inhibition of the synthesis of alpha-globin was observed only in the cells transformed with the plasmids with antisense 6 ...	1992	1295698
enzymatic analysis of oligonucleotides containing cyclobutane pyrimidine photodimers with a cleaved intradimer phosphodiester linkage.	our recent studies indicate that enzymatic hydrolysis of the intradimer phosphodiester linkage constitutes an early reaction in processing uv light-induced cis-syn-cyclobutane pyrimidine dimers in cultured human fibroblasts. before characterizing the resultant modified dimer sites in cellular dna, it is necessary to establish experimental conditions that can distinguish backbone-nicked from intact dimers. we thus constructed a model substrate, i.e. p(dt) 10 <> p(dt)10 containing a dimer with a r ...	1992	1331055
consequences of 6-thioguanine incorporation into dna on polymerase, ligase, and endonuclease reactions.	the incorporation of 6-thioguanine (s6g) in place of guanine proceeds readily in dna synthesis reactions catalyzed by mammalian and bacterial polymerases. this report summarizes the consequences of such incorporation studied to date. s6g was incorporated into one strand of a defined m13mp18 phage sequence in a (+)reaction catalyzed by the klenow fragment of escherichia coli dna polymerase i. after denaturation of the newly synthesized strand (containing s6g) and annealing with a reverse (-) 32p- ...	1992	1331762
intraoperative monitoring of tibialis anterior muscle motor evoked responses to transcranial electrical stimulation during partial neuromuscular blockade.	we studied the feasibility of recording motor evoked responses to transcranial electrical stimulation (tce-mers) during partial neuromuscular blockade (nmb). in 11 patients, compound muscle action potentials were recorded from the tibialis anterior muscle in response to transcranial electrical stimulation during various levels of vecuronium-induced nmb. the level of nmb was assessed by accelerometry of the adductor pollicis muscle after train-of-four stimulation of the ulnar nerve. the compound ...	2005	1356320
rapd analysis of campylobacter isolates: dna fingerprinting without the need to purify dna.	a method was developed to obtain reproducible dna fingerprints from campylobacter by pcr-based amplification, without the need to isolate total dna. randomly amplified polymorphic dna (rapd) profiles were generated with three randomly designed 10-mers, using each separately as an amplification primer. a range of c. jejuni serotypes could be typed by rapd analysis. depending on the primer, the analysis of rapd profiles resulted in different levels of discrimination between the strains. clear corr ...	1992	1368370
functional analysis of the mouse alpha-fetoprotein enhancers and their subfragments in primary mouse hepatocyte cultures.	we have compared the activities of mouse alpha-fetoprotein (afp) enhancers i, ii, and iii with their minimal enhancer fragments (mers) i, ii, and iii and with the entire 7-kilobase pair enhancer domain by transient expression assay in primary fetal mouse liver cells. the level of expression directed by the afp promoter [p(-1009)afpcat] alone is stimulated at least 10-fold by the entire afp enhancer domain (-1009 to -6983). enhancer i can drive the level of chloramphenicol acetyltransferase activ ...	1992	1375227
antisense oligodeoxynucleotides to the cystic fibrosis transmembrane conductance regulator inhibit camp-activated but not calcium-activated chloride currents.	phosphorylation of the cystic fibrosis transmembrane conductance regulator (cftr) by camp-dependent protein kinase leads to chloride flux in epithelial cells. is cftr also required for the calcium-dependent activation of chloride channels? we used antisense oligodeoxynucleotides to cftr to reduce the expression of cftr in colonic and tracheal epithelial cells. the antisense oligomers were a pair of adjacent 18-mers complementary to nucleotides 1-18 and 19-36 of cftr mrna. sense and misantisense ...	1992	1379720
template. phosphorothioate oligonucleotides duplexes as inhibitors of hiv-1 reverse transcriptase.	we have investigated the interaction between a number of 14 mers phosphorothioate oligonucleotides and hiv-1 reverse transcriptase. two methods were used to measure the affinity of the analogs for the enzyme. in the first, the oligonucleotide or its duplex with poly(rl) were used as inhibitors of the enzyme using poly(ra).(dt)14 as template primer. in the second, the oligonucleotides or their duplexes were used to displace a fluorescent template primer complex of known affinity from its binding ...	1992	1380799
differential mapping of fc gamma-binding and monoclonal antibody-reactive epitopes on ge, the fc gamma-binding glycoprotein of herpes simplex virus type 1.	the entire 396 residue extracellular sequence of ge the hsv-1 fc gamma-binding glycoprotein has been studied to determine epitopes binding to two mab ii-481 and 88s previously demonstrated to react with ge at or near the fc gamma-binding regions. overlapping 7-mers constructed from the established sequence were tested with mab ii-481 and 88s along with their fab fragments. control mab of the same igg 2b subclass as well as whole rabbit and human igg and fc were also tested for binding to overlap ...	1992	1382102
lipofectin enhances cellular uptake of antisense dna while inhibiting tumor cell growth.	a natural dna oligomer (15-mer) was synthesized with a sequence complementary to the translation initiation codon region of the human tgf-alpha mrna and mixed with lipofectin to form unilamellar complexes. it was found that tumor cell growth was inhibited when hct116 cells were treated with lipofectin-dna oligomer complexes or with lipofectin alone. uptake of 32p-labeled 15-mers into colon tumor cells was compared in the presence and absence of lipofectin. the amount of labeled oligomer found in ...	1992	1422086
opening of the replication origin of escherichia coli by dnaa protein with protein hu or ihf.	opening of the three tandem repeats of a 13-mer in the replication origin (oric) of escherichia coli is a prime event in the replication in vitro of minichromosomes (bramhill, d., and kornberg, a. (1988) cell 54, 915-918). dnaa, the initiator protein, requires protein hu or ihf, along with a millimolar level of atp and negative superhelical density in the plasmid to open this region. the extent of opening, as judged by cleavage by a single-strand-specific endonuclease (i.e. p1 nuclease), correla ...	1992	1429655
opposed actions of regulatory proteins, dnaa and icia, in opening the replication origin of escherichia coli.	the opening of the three tandem 13-mers (iterons) in the replication origin (oric) of escherichia coli by dnaa protein, assisted by protein hu or ihf (hwang, d. s., and kornberg, a. (1992) j. biol. chem. 267, 23083-23086), represents an essential early stage in the initiation of chromosomal replication (bramhill, d., and kornberg, a. (1988) cell 54, 915-918). we now show by mutational alterations of the 13-mer region that oric function, both in vitro and in vivo, requires at-richness in the left ...	1992	1429656
microtubule-associated protein tau is required for axonal neurite elaboration by neuroblastoma cells.	nb2a/d1 neuroblastoma cells constitutively express multiple isoforms of the microtubule-associated protein tau and incorporate this protein into the axonal neurites elaborated during serum deprivation. to examine whether or not tau played an essential role in axonal outgrowth, cells cultured in serum-free medium were treated at 24 h intervals with antisense- and sense-oriented cdna oligonucleotides (25 or 36 mers that span or are upstream of tau initiation codon) and were simultaneously serum de ...	1992	1433385
an oligomer complementary to the 5' end region of mdr1 gene decreases resistance to doxorubicin of human adenocarcinoma-resistant cells.	acquired resistance to doxorubicin and other anti-cancer drugs is generally dependent on gene amplification of a specific nucleotide sequence, the mdr1 gene. verapamil, cyclosporin and other drugs have been used to circumvent the resistance in experimental models in vitro and/or in vivo. we have attempted to reverse the mdr phenotype by treating human adenocarcinoma resistant cells with 20 mers of synthetic unmodified oligodeoxynucleotide mdr1 antisenses. five odns towards different mrna regions ...	1992	1444203
use of specific oligonucleotides for direct enumeration of listeria monocytogenes in food samples by colony hybridization and rapid detection by pcr.	two 18-mer oligonucleotides derived from the sequence of hly, the gene coding for listeriolysin o, were shown to be specific for listeria monocytogenes in the genus listeria in colony hybridization tests. the oligonucleotides did not hybridize with any of the bacterial species found in food and co-isolated with listeria on selective media. they were used in colony hybridization tests for enumeration of l. monocytogenes present in food samples after direct plating on selective media plates. in ad ...	1992	1448613
sequencing by hybridization: towards an automated sequencing of one million m13 clones arrayed on membranes.	an immediately applicable variant of the sequencing by hybridization (sbh) method is under development with the capacity to determine up to 100 million base pairs per year. the proposed method comprises six steps: (i) arraying genomic or cdna m13 clones in 864-well plates (wells of 2 mm); (ii) preparation of dna samples for spotting by growth of the m13 clones or by polymerase chain reaction (pcr) of the inserts using standard 96-well plates, or plates having as many as 864 correspondingly small ...	1992	1451694
genotypic identification and characterization of species and strains within the genus candida by using random amplified polymorphic dna.	random amplified polymorphic dna (rapd) was used to better characterize the genotypic relatedness among medically important candida species. by using short oligomer primers (10-mers) with arbitrarily chosen sequences in the polymerase chain reaction, distinctive and reproducible sets of polymerase chain reaction products were observed for isolates of c. albicans, c. lusitaniae, c. tropicalis, and torulopsis (candida) glabrata. the rapd analysis differentiated a physiologically homogeneous panel ...	1992	1452710
specific inhibition of human immunodeficiency virus type 1 replication by antisense oligonucleotides: an in vitro model for treatment.	we have developed a culture system, simulating in vivo conditions of human immunodeficiency virus type 1 (hiv-1) infection, to evaluate the long-term efficacy of antisense oligonucleotide treatment. five oligonucleotide phosphorothioates (28-mers), complementary to different regions of hiv-1 rna, blocked replication of the virus in a sequence-specific manner at 1 microm concentration. variations in antiviral activity were seen among the different oligonucleotides, revealing an effect of target s ...	1992	1454800
studies on antisense inhibition of translation in vitro. anomalies and re-evaluation.	experiments were carried out to better characterize antisense control of translation. results in an e. coli system confirmed specific inhibition of poly(u) translation. at low concentrations, certain homopolymers (including poly(ra)) stimulated translation. oligo(da(n)) was inhibitory at n less than or equal to 8. translation of globin mrna in reticulocyte lysates indicated that ssdna 15-mers targeted at beta-globin mrna inhibited both alpha- and beta-globin production. sequences targeted immedi ...	1992	1516711
in vivo primary induction of virus-specific ctl by immunization with 9-mer synthetic peptides.	a primary cytotoxic t lymphocyte (ctl) response in vivo requires antigen presentation by cytosolic processing and can not in general be obtained by vaccination with soluble proteins. in the present work we have found that vaccination of mice with pre-processed synthetic peptides, corresponding to endogenous 9-mers produced in influenza a virus-infected cells, resulted in strong primary ctl responses. the generated ctl efficiently killed virus-infected target cells with preference for viral strai ...	1992	1517589
rna polymerases react differently at d(apg) and d(gpg) adducts in dna modified by cis-diamminedichloroplatinum(ii).	two duplexes (20-mers) were constructed containing either a single cis-[pt(nh3)2[d(gpg)]] or cis-[pt(nh3)2[d(apg)]] intrastrand cross-link, the major dna adducts of the antitumor drug cis-diamminedichloroplatinum(ii). these synthetic duplexes were multimerized and the resultant polymers used as templates in single-step addition reactions of condensation of a single nucleoside triphosphate substrate to a dinucleotide primer (abortive elongation reaction) catalyzed by prokaryotic or eukaryotic rna ...	1992	1536834
inhibition of t4 dna ligase activity by (+)-cc-1065: demonstration of the importance of the stiffening and winding effects of (+)-cc-1065 on dna.	non-denaturing gel electrophoresis analysis demonstrates that the stiffening and winding effects of (+)-cc-1065 produce unusual proximal and distal inhibition of t4 dna ligase-catalysed ligation of covalently modified dna. (+)-cc-1065 is a potent antitumor antibiotic produced by streptomyces zelensis. this drug selectively bonds through n3 of adenine in dna and lies in the minor groove of dna, reacting in a highly sequence-selective manner. previous studies (lee et al., 1991) have shown that (+) ...	1992	1543525
structure of the dnaa and dnaa-box region in the mycoplasma capricolum chromosome: conservation and variations in the course of evolution.	we have previously shown that the dnaa gene and the dnaa-box region were conserved in bacteria representative of all three major branches of the eubacterial phylogenic tree: high g + c gram+, low-g + c gram+ and gram-. in the present work, we determined the structure of the dnaa region of mycoplasma capricolum and found that the dnaa gene and at least two other genes, rpmh and dnan, were conserved in this bacterium. an unusually high level of amino acid (aa) substitutions was observed in m. capr ...	1992	1544573
effects of propofol, etomidate, midazolam, and fentanyl on motor evoked responses to transcranial electrical or magnetic stimulation in humans.	the effects of propofol, etomidate, midazolam, and fentanyl on motor evoked responses to transcranial stimulation (tc-mers) were studied in five healthy human volunteers. each subject, in four separate sessions, received intravenous bolus doses of propofol 2 mg.kg-1, etomidate 0.3 mg.kg-1, midazolam 0.05 mg.kg-1, and fentanyl 3 micrograms.kg-1. electrical tc-mers (tce-mers) were elicited with anodal stimuli of 500-700 v. magnetic tc-mers (tcmag-mers) were elicited using a cadwell mes-10 magnetic ...	1992	1550274
hla-a2 molecules in an antigen-processing mutant cell contain signal sequence-derived peptides.	the mutant human cell line t2 is defective in antigen presentation in the context of class i major histocompatibility complex (mhc) molecules, and also in that transfected t2 cells show poor surface expression of exogenous human class i (hla) alleles. both defects are thought to lie in the transport of antigenic peptides derived from cytosolic proteins into the endoplasmic reticulum (er), as peptide-deficient class i molecules might be expected to be either unstable or retained in the er. the pr ...	1992	1557127
a pcr-mediated gene synthesis strategy involving the assembly of oligonucleotides representing only one of the strands.	a modification of pcr-mediated gene synthesis strategy is introduced. this modification enables the synthesis of a gene from oligonucleotides comprising only one of the two strands. bridging oligonucleotides (approximately 20-mers in length) complementary to the junctions of template strand oligonucleotides and two outer primers are also needed for pcr. a two-step pcr containing a first step of 10 cycles, followed by a second step of 20 cycles, differing only in the annealing conditions was used ...	1992	1571150
a novel blocker-pcr method for detection of rare mutant alleles in the presence of an excess amount of normal dna.	a novel polymerase chain reaction method was developed to preferentially amplify a segment of dna containing a base substitution mutation. this technique uses a pair of dideoxynucleotide-labeled oligonucleotides (18 mers) of normal sequences as blockers located between the two primers. by virtue of a subtle difference in the melting temperature between the blocker-normal dna and blocker-mutant dna hybrids, the method allows preferential amplification of the mutant dna. we used the human n-ras ge ...	1992	1598207
uranyl photofootprinting of triple helical dna.	two triple helix structures (15-mers containing only t.a-t triplets or containing mixed t.a-t and c.g-c triplets) have been studied by uranyl mediated dna photocleavage to probe the accessibility of the phosphates of the dna backbone. whereas the phosphates of the pyrimidine strand are at least as accessible as in double stranded dna, in the phosphates of the purine strand are partly shielded and more so at the 5'-end of the strand. with the homo a/t target increased cleavage is observed towards ...	1992	1614860
the immobiline family: from "vacuum" to "plenum" chemistry.	we list here a total of 17 acrylamido acids and bases as potential buffers and titrants for isoelectric focusing separations in immobilized ph gradients. the chemistry of these compounds is reviewed and general guidelines are given for their proper use. in particular, it is shown that the most delicate compounds are the basic species, since they can undergo several degradation pathways, including: (i) spontaneous hydrolysis to acrylic acid and a diamine; (ii) spontaneous autopolymerization to ol ...	1992	1628597
identification of active peptide sequences in the carboxyl-terminal cell binding domain of human thrombospondin-1.	thrombospondin (ts) mediates attachment, spreading, and motility of several cell types through at least four cell binding domains: the amino-terminal heparin binding domain, the type i repeats containing the csvtcg sequence, the rgda sequence in the last of the type iii calcium binding repeats and the carboxyl-terminal cell or platelet binding domain (cbd). the attachment of human melanoma cells (g361) to the cooh-terminal domain is independent of the rgda sequence and is inhibited by the monocl ...	1992	1644809
effects of 2-chloroadenine substitution in dna on restriction endonuclease cleavage reactions.	the purine analog, 2-chloro-2'-deoxyadenosine triphosphate (cldatp), was incorporated enzymatically in place of datp into the minus strand of m13mp18 duplex dna. its effect on protein-dna interactions was assessed by determining the amount of dna cleavage by type ii restriction endonucleases. substitution of chloroadenine (ciade) for adenine (ade) in dna appreciably decreased the amount and rate of dna cleavage of the minus strand when the analog was situated within the appropriate endonuclease ...	1991	1647525
preparation and characterization of spin-labeled oligonucleotides for dna hybridization.	sequence-specific spin-labeled oligodeoxynucleotides with conformation-sensitive electron paramagnetic resonance (epr) signals are synthesized and examined as solution-phase nucleic acid hybridization probes. either a proxyl or tempo ring linked to the c(5) position of deoxyuridine (du) by a nonrigid two-atom methylamino tether is incorporated within 15-mers by phosphotriester chemistry yielding stable spin-labeled probes with distinctive epr specific activity (aepr) values. the aepr is greater ...	2013	1651116
identification of skeletal muscle protein-tyrosine phosphatases by amplification of conserved cdna sequences.	specific protein-tyrosine phosphatase (ptpase) enzymes that regulate signal transduction by the insulin receptor in target tissues have not been identified. we evaluated the expression of ptpase homologs in skeletal muscle since this tissue is the major site of insulin-mediated glucose disposal in vivo. a rat skeletal muscle cdna pool was prepared with a set of degenerate oligonucleotide primers and ptpase cdna sequences were amplified using pairs of "guess-mers" that were deduced from highly co ...	1991	1651716
purification, properties, and mutagenesis of poliovirus 3c protease.	poliovirus protease 3c, type 1 mahoney strain, was expressed in escherichia coli under phage t7 promoter control and purified to homogeneity from resolubilized inclusion bodies. the renatured protein was as enzymatically active as the protease found in the soluble portion of the bacterial lysate. proteolytic activity was assayed using as substrate either [35s]methionine-labeled recombinant poliovirus proteins 2c3ab or a truncated version of 3abc, or synthetic peptide 16-mers corresponding to the ...	1991	1656583
mechanism of cellular uptake of modified oligodeoxynucleotides containing methylphosphonate linkages.	the cellular uptake and intracellular distribution of methylphosphonate oligonucleotides (15 mers) has been examined using both 32p labeled and fluorescent labeled oligonucleotides. the cellular uptake process for methylphosphonate oligonucleotides is highly temperature dependent, with a major increase in uptake occurring between 15 and 20 degrees c. most of the label which becomes cell associated at 37 degrees c cannot be removed by acid washing or trypsinization and thus seems to be within the ...	1991	1658734
intermediates in the chaperonin-assisted refolding of rhodanese are trapped at low temperature and show a small stoichiometry.	in vitro refolding of the urea-unfolded, monomeric, mitochondrial enzyme rhodanese (thiosulfate sulfur-transferase; ec 2.8.1.1) is facilitated by the chaperonin proteins cpn60 and cpn10 from escherichia coli at 37 degrees c, but the refolding is strongly inhibited at 10 degrees c. in contrast, the unassisted refolding of rhodanese is efficient at 10 degrees c, but the refolding efficiency decreases as the temperature is raised. these observations provided two measures of the cpn60-rhodanese comp ...	1991	1680127
inhibition of the p66/p51 form of human immunodeficiency virus reverse transcriptase by trna(lys).	human immunodeficiency virus (hiv) reverse transcriptase (rt) uses host trna(lys) partially annealed to the primer binding site (pbs) as primer for the initiation of cdna synthesis. when assaying cdna synthesis with a template-primer complex formed by an rna fragment carrying the pbs site and bovine trna(lys) we noticed that an excess of primer trna inhibited strongly the dna polymerase activity of a recombinant hiv rt (p66-p51 heterodimeric form) produced in transformed yeast cells. the same in ...	1990	1689823
identification and characterization of t helper cell epitopes of the major outer membrane protein of chlamydia trachomatis.	chlamydia trachomatis serovars a, b, and c are the causative agents of trachoma, the world's leading cause of preventable blindness. immunoprophylaxis is a possible approach to control trachoma. the chlamydial major outer membrane protein (momp) is thought to play an important role in the development of protective immunity against chlamydial infection, and is therefore considered to be a promising candidate antigen in the development of a trachoma vaccine. much effort has been focused on the mol ...	1990	1694217
herpes simplex virus type 1-specific immunity induced by peptides corresponding to an antigenic site of glycoprotein b.	herpes simplex virus (hsv) envelope glycoproteins are the prime targets of adaptive antiviral immunity. previous investigation identified a protective, neutralizing, glycoprotein b1 (gb-1)-reactive monoclonal antibody (mab b6) and localized the linear epitope recognized by the mab to residue 84 of gb-1. three overlapping peptides (two 20-mers and one 18-mer), together spanning amino acids 63 to 110 of the wild-type sequence of gb-1, were synthesized and analyzed for their ability to stimulate im ...	1990	1698994
[effective synthesis of oligo(poly)deoxyribonucleotides using an h-phosphonate method in plastic microcolumn].	a facile technique of manual oligonucleotide synthesis via h-phosphonate approach is developed. syntheses carried out in pipette tips with siliconised glasswool filters take 3-3.5 min per cycle with 97-98% yields per condensation. the method was used to synthesize 12-55-mers: t7 and pl promoter regions, gene of the signal peptide of the e. coli ompa protein, oligonucleotides coding for amino acid sequences 94-105 of pres1- and 133-143 of pres2-regions of hepatitis b virus, hybridisation probes, ...	1990	1700717
differential effects of lecithin and cholesterol on the immunoreactivity and conformation of apolipoprotein a-i in high density lipoproteins.	recently identified epitopes in apoa-i define a distinct n-terminal region with a complex tertiary structure, characterized by multiple discontinuous epitopes. other epitopes are constituted of short domains centered either on beta-turns or random coils or on the 22-mer amphipathic alpha-helices (marcel, y. l., provost, p. r., koa, h., raffaï, e., vu dac, n., fruchart, j.-c., and rassart, e. (1991) j. biol. chem. 266, 3644-3653). the compared immunoreactivity of seven epitopes studies here in re ...	1991	1709164
identification of two t-cell epitopes on the candidate epstein-barr virus vaccine glycoprotein gp340 recognized by cd4+ t-cell clones.	current efforts to develop an epstein-barr virus subunit vaccine are based on the major envelope glycoprotein gp340. given the central role of cd4+ t cells in regulating immune responses to subunit vaccine antigens, the present study has begun the work of identifying linear epitopes which are recognized by human cd4+ t cells within the 907-amino-acid sequence of gp340. a panel of gp340-specific cd4+ t-cell clones from an epstein-barr virus-immune donor were first assayed for their proliferative ...	1991	1710291
duplex stabilities of phosphorothioate, methylphosphonate, and rna analogs of two dna 14-mers.	the duplex stabilities of various phosphorothioate, methylphosphonate, rna and 2'-och3 rna analogs of two self-complementary dna 14-mers are compared. phosphorothioate and/or methylphosphonate analogs of the two sequences d(taattaattaatta) [d1] and d(tagctaattagcta) [d2] differ in the number, position, or chirality (at the 5' terminal linkage) of the modified phosphates. phosphorothioate derivatives of d1 are found to be less destabilized when the linkage modified is between adenines rather than ...	1991	1711677
a cluster of continuous antigenic structures in the transmembrane protein of hiv-1: individual patterns of reactivity in human sera.	we investigated the antigenicity of a highly conserved region in the transmembrane protein of the human immunodeficiency virus type 1 (hiv-1). in order to identify antigenically important residues, amino-acid sequences of synthetic peptides representing this region were varied systematically: single residues were omitted from the sequence of hiv-env 583-599; threonines were substituted for pairs of residues in hiv-env 581-599; the sequences of heptadeca-peptides were shifted by single residues. ...	1991	1713646
identification of an immunodominant b cell epitope on the hepatitis c virus nonstructural region defined by human monoclonal antibodies.	several ebv-transformed b cell lines (bcl) were obtained from two patients with chronic hepatitis c virus (hcv) infection that secreted igg class antibodies to the hcv nonstructural ag c100-3. two cloned bcl, derived from the same parental line, generated stable cloned lines that secreted up to 20 mg/liter of specific igg1(kappa). supernatants from oligoclonal and cloned bcl were also analyzed by immunoblot and all strongly reacted with recombinant polypeptides derived from the putative ns4 regi ...	1991	1717573
epitope mapping of snake venom phospholipases a2 with pseudexin monoclonal antibodies.	fifteen different monoclonal antibodies, developed against a pseudexin a, b, and c mixture, were screened for linear epitope recognition. peptides (9-mers) spanning pseudexin b were synthesized on alanine-derivatized polyethylene pins and subsequently probed with antibody. four antibodies recognized linear epitopes of pseudexin a, pseudexin b, and also nonidentical sequences found in other phospholipases a2 (pla2s) as determined by enzyme-linked immunosorbent assays. three antibodies recognized ...	1991	1718309
structure and assembly of the escherichia coli transcription termination factor rho and its interaction with rna. i. cryoelectron microscopic studies.	cryoelectron microscopy has been used to visualize the escherichia coli transcription termination protein rho in a vitreously frozen state, without the use of strains, fixatives or other chemical perturbants. in the absence of rna cofactor, a variety of structures are observed, reflecting the heterogeneity of complexes formed by rho at protein concentrations near the physiological range (3 to 10 microm). one of the most common structural motifs we see is a six-membered ring of rho subunits (pres ...	1991	1719215
identification of antibody epitopes within the cb-11 peptide of type ii collagen. ii. computer modelling studies of peptides and the interpretation of epitope scanning results.	computer modelling techniques were used to investigate the structure of 8-mers from the cb-11 peptide of bovine type ii collagen which were recognised by sera from rats which had previously been injected with bovine type ii collage. it was discovered that all the hydrophobic peptides recognised by the rat sera were predicted to have collagenous-like secondary structures. the primary structure of the 8-mers which were recognised was also compared against the sequences in the owl protein sequence ...	1991	1721847
epitope mapping studies of snake venom phospholipase a2 using monoclonal antibodies.	fifteen different monoclonal antibodies developed against pseudexin, a snake venom phospholipase a2 with presynaptic neurotoxicity, were screened for linear epitope recognition. peptides (9-mers) spanning pseudexin were synthesized by using alanine-derivatized polyethylene pins and subsequently probed with antibody. four antibodies bound to toxin peptides and were detected with an enzyme-linked immunosorbent assay. three of the bound antibodies recognized a site important in calcium binding and ...	1991	1725235
icia protein, a specific inhibitor of initiation of escherichia coli chromosomal replication.	specific binding of icia protein to the 13-mers in the origin of a minichromosome (oric) inhibits initiation of replication in vitro by blocking the opening of this region effected by the initiator dnaa protein (hwang, d.s., and kornberg, a. (1990) cell 63, 325-331). isolation of the icia gene (thöny, b., hwang, d.s., fradkin, l., and kornberg, a. (1991) proc. natl. acad. sci. u.s.a. 88, 4066-4070) has made possible the construction of an icia-overproducing strain, which in turn has simplified t ...	1992	1733927
transcription in vivo within the replication origin of the escherichia coli chromosome: a mechanism for activating initiation of replication.	within the replication origin, oric, of the escherichia coli chromosome, novel in vivo transcripts were detected which proceeded rightward and whose production was activated by dnaa protein. in contrast, dnaa protein repressed the previously described ori-l leftward transcription. the former should introduce negative supercoiling, and the latter positive supercoiling, into the 13-mers. the effects of transcription on the initiation of replication were also investigated by making constructs with ...	1992	1736090
site-specific interaction of the antitumor antibiotic dynemicin with branched dna molecules.	a specific interaction of stable branched dna molecules with the antitumor antibiotic dynemicin is reported. dynemicin contains an anthraquinone and an enediyne unit, and belongs to the family of enediyne antitumor agents. dna strand scission by dynemicin appears to involve interaction of the anthraquinone core with dna and release of a phenyl diradical from the enediyne core that can abstract hydrogen atoms from the sugar phosphate backbone of dna. the cleavage patterns of each labeled strand i ...	1991	1741963
analysis of salmonella typhimurium hisd3052 revertants: the use of oligodeoxyribonucleotide colony hybridization, pcr, and direct sequencing in mutational analysis.	a rapid method for determining the dna sequences of salmonella typhimurium hisd3052 revertants is presented. dna colony hybridization was used to analyze revertants previously studied by isono and yourno [proc natl acad sci usa 71:1612-1617, 1974]. synthetic oligodeoxyribonucleotide probes (18-mers) were able to distinguish sequences that differed by a single base pair. mutant his sequences not identified by probing analysis were amplified using polymerase chain reaction (pcr) and directly seque ...	1991	1748083
[suppression of translation in vitro of the mrna of the m1 protein of influenza virus using antisense oligonucleotides].	effect of antisense oligonucleotides on the in vitro translation of the influenza virus m1 protein mrna was investigated. the most efficient arrest of mrna translation was achieved by simultaneous action of two or three oligonucleotides (14-16-mers) complementary to the juxtaposed sequences in the 5'-terminus of the molecule around and upstream of the initiation codon.	1991	1753959
a method for dna sequencing by hybridization with oligonucleotide matrix.	a new technique of dna sequencing by hybridization with oligonucleotide matrix (shom) which could also be applied for dna mapping and fingerprinting, mutant diagnostics, etc., has been tested in model experiments. a dot matrix was prepared which contained 9 overlapping octanucleotides (8-mers) complementary to a common 17-mer. each of the 8-mers was immobilized as individual dot in thin layer of polyacrylamide gel fixed on a glass plate. the matrix was hybridized with the 32p-labeled 17-mer and ...	1991	1768861
hybridization methods for dna sequencing.	i have conducted a general analysis of the practicability of using oligonucleotide hybridization to sequence dna. any dna sequence may be sequenced by hybridization with a complete panel of oligonucleotides. however, sequencing dna segments over 2 kb long requires an unrealistic number of hybridization reactions. the optimal protocol is to hybridize 7-mer or 8-mer mixed oligonucleotide probes to immobilized dna fragments 80 bp long: should this prove impractical, hybridization of labeled 270-bp ...	1991	1769648
[oncogene-directed mutagenesis in vivo. polyalkylating derivatives of short single-stranded polynucleotides, complementary e1-adeno-oncogene, in the normalization of adenovirus-transformed rodent cell lines].	polyalkylating derivatives of single-stranded polynucleotides (30-200-mers) complementary to the long e1 oncogene sequences of simian adenovirus sa7 cause inherited normalization of sh2 and g11 cells transformed with adenovirus sa7; certain deletions in the integrated proviral e1a oncogene were observed in several cases during this process. the transformed cells are indifferent to reagents noncomplementary to the e1 region. thus polyalkylating derivatives of single-stranded 30-200-mers act as ad ...	2006	1795710
retention behaviour of a template-assembled synthetic protein and its amphiphilic building blocks on reversed-phase columns.	the retention behaviour of a six-helix bundle template-assembled synthetic protein (tasp) molecule and its amphiphilic building blocks was investigated. the tasp consists of a circular template, cyclo(1-12)[kg]6, and six identical potentially alpha-helical peptides of the sequence klalklalkalklalkla. as an alpha-helix, this peptide is amphiliphilic along the axis of its helix. based on this sequence, the retention times of a set of acetylated peptides containing from seven to twenty amino acids ...	1991	1806554
co-operative autoregulation of a replication protein gene.	in this work we present the localization and characterization of the repl promoter (prepl) and show aspects of the regulation. comparison of prepl with other autoregulated replication protein gene promoters revealed similarities, but prepl differs from some of these characterized promoters in not being regulated by the heat-shock rna polymerase. primer extension analysis showed that prepl is contained within five helically aligned 18 base pair repeats, or 18-mers of the previously defined minima ...	1991	1809840
non-sequence-specific inhibition of transferrin receptor expression in hl-60 leukemia cells by phosphorothioate oligodeoxynucleotides.	a series of phosphodiester and phosphorothioate antisense oligodeoxynucleotides were synthesized against the human transferrin receptor (tfr). the phosphorothioate analogs exhibited marked biologic efficacy in culture, as assessed by inhibition of surface tfr content and hl-60 cell growth, whereas their unmodified phosphodiester counterparts were ineffective. phosphorothioate oligodeoxynucleotides were more resistant to hydrolysis by serum and cellular nucleases and were more readily taken up by ...	1991	1821654
low concentrations of isoflurane abolish motor evoked responses to transcranial electrical stimulation during nitrous oxide/opioid anesthesia in humans.	to study the feasibility of noninvasive monitoring of motor pathways in anesthetized patients, we evaluated the effect of isoflurane on motor evoked responses to constant-voltage transcranial electrical stimulation (tce-mers). reproducible tce-mers were recordable from the tibialis anterior muscle during nitrous oxide/opioid anesthesia in 11 patients. before the introduction of isoflurane, tce-mer onset latency was 30.8 +/- 1.9 ms, and amplitude ranged from 19 microv to 2.6 mv (median, 209 micro ...	1991	1832825
oligonucleotides antisense to the interleukin 1 receptor mrna block the effects of interleukin 1 in cultured murine and human fibroblasts and in mice.	phosphodiester and phosphorothioate oligodeoxynucleotides (18 mers) were constructed antisense to sequences of the recently cloned murine and human il-1 receptors. murine antisense oligonucleotides inhibited il-1-stimulated pge2 synthesis by murine fibroblasts in culture in a time (days) and concentration-dependent (3 microm-30 microm) fashion. murine sense oligonucleotide and an oligonucleotide antisense to human il-1 receptor were without effect. moreover, murine antisense oligonucleotides did ...	1991	1833422
complementary directed modifications of nucleic acids and oncogene-directed mutagenesis in vivo.	high reactivity of the polyalkylating ss oligomers that were sense or antisense 30-200-mers containing sequences complementary to e1 oncogenes of simian adenovirus sa7 and one alkylating residue -ch2ch2n(c2h5oh) (ch2)3n(ph-p-ch2oh)ch2ch2cl per each 25 bases of oligomers was demonstrated in vitro by alkylation of ss dna of recombinant m 13 mp8e1 and mp9e1 phages with inserted e1 sequences of adenovirus oncogene and then by followed complete and selective elimination of e1 sequences from recombina ...	1991	1841269
analysis of the molecular mimicry between hla-b27 and a bacterial ompa protein using synthetic peptides.	in spite of a lack of sequence 'homology' between hla-b27 and the bacterial ompa outer membrane proteins, they both react with the ye-2 monoclonal anti-hla-b27 antibody. the ye-2 antibody also reacted positively in elisa with a synthetic peptide derived from the segment spanning residues 63-84 of b*2705. the critical peptide residues were determined by testing first with overlapping peptides, followed by a replacement set made according to the determined epitope. the results were compared with t ...	1991	1893633
a barbiturate-regulated protein binding to a common sequence in the cytochrome p450 genes of rodents and bacteria.	analyses of the 5' regulatory sequences of genes encoding barbiturate-inducible cytochromes p450bm-1 and p450bm-3 from bacillus megaterium and of the 5' sequences of genes for barbiturate-inducible p450b and p450e of the rat revealed a string of 17 base pairs in each of the genes that shared a high degree of sequence identity. labeled oligonucleotide probes of each of these four sequences were tested in gel retardation assays with protein obtained from b. megaterium grown either in the presence ...	1991	1902228
dna sequence dependence of guanine-o6 alkylation by the n-nitroso carcinogens n-methyl- and n-ethyl-n-nitrosourea.	after intracellular in vitro exposure to the mutagenic and carcinogenic n-nitroso compounds n-methyl-n-nitrosourea (menu) or n-ethyl-n-nitrosourea (etnu), respectively, the average relative amounts of the premutational lesion o6-alkylguanine represent about 6% and 8% of all alkylation products formed in genomic dna. at the level of individual dna molecules guanine-o6 alkylation does not occur at random; rather, the probability of a substitution reaction at the nucleophilic o6 atom is influenced ...	2013	1944330
polymorphism in n-2-acetylaminofluorene induced dna structure as revealed by dnase i footprinting.	in this paper, we have constructed double stranded helices (60-mers) containing a single n-2-acetylaminofluorene (-aaf) adduct covalently bound to one of the three guanine residues of the narl site (g1g2cg3cc). this sequence was identified as a strong frameshift mutation hot spot for many carcinogens that bind to the c8 position of guanine. using dnase i as a probe for dna conformation we show i) that the average size of the helix deformation extends over 3 to 5 base pairs in both directions fro ...	1991	1945836
solid phase synthesis of oligodeoxyribonucleoside phosphorodithioates from thiophosphoramidites.	oligonucleoside phosphorodithioates 1 are modified dna sequences with potential use as antisense oligonucleotides. the preparation of up to 20-mers containing all four bases by solid phase synthesis is described, with details on the preparation of the four monomer units (protected nucleoside thiophosphoramidites 2), the conditions used for the assembly of the strands with up to 19 phosphorodithioate linkages, and the purification and characterisation of the products. full-length homogeneity of h ...	1991	1945874
the properdin-like type i repeats of human thrombospondin contain a cell attachment site.	thrombospondin (ts) is a modular adhesive glycoprotein that contains three domains previously implicated in the attachment of cells to ts. these include the amino-terminal heparin-binding domain, the carboxy terminal cell or platelet-binding domain, and an rgda sequence of ts. we have characterized a mab against human ts, designated a4.1, which inhibits the attachment of human melanoma cells (g361) to ts. the epitope for a4.1 lies within the amino terminal half of the central stalklike region of ...	1991	1999454
ecdysterone regulatory elements function as both transcriptional activators and repressors.	a synthetic, 23-bp ecdysterone regulatory element (ecre), derived from the upstream region of the drosophila melanogaster hsp27 gene, was inserted adjacent to the herpes simplex virus thymidine kinase promoter fused to a bacterial gene for chloramphenicol acetyltransferase (cat). hybrid constructs were transfected into drosophila s3 cells and assayed for ecdysterone-inducible cat expression. in the absence of ecdysterone a tandem pair of ecres repressed the high constitutive level of cat activit ...	1991	2005885
intramolecular triplex formation of the purine.purine.pyrimidine type.	six octadecamers with hairpin motifs have been synthesized and investigated for possible intramolecular triplex formation. electrophoretic, hypochromic, and cd evidence suggest that d(cccctttggggtttgggg) and d(ggggtttggggtttcccc) can form g.g.c intramolecular triplexes via double hairpin formation in neutral solutions, presumably with the terminal g tract folding back along the groove of the hairpin duplex. in contrast, d(ggggtttcccctttgggg) and the three corresponding 18-mers containing one g a ...	1991	2021637
icia, an escherichia coli gene encoding a specific inhibitor of chromosomal initiation of replication in vitro.	the gene encoding the protein that binds the three 13-mers in the origin (oric) of escherichia coli to block initiation of replication in vitro has been cloned, sequenced, and overexpressed. the gene possesses an open reading frame for 297 amino acids (mass of 33,471 da). the protein has a motif for dna-binding (helix-turn-helix) and has homology to a diverse set of prokaryotic regulatory proteins, known as the lysr family. the protein, previously referred to as the 33-kda protein, has been name ...	1991	2034653
influence of the incorporation of (s)-9-(3,4-dihydroxybutyl)adenine on the enzymatic stability and base-pairing properties of oligodeoxynucleotides.	(s)-9-(3,4-dihydroxybutyl)adenine was used at several positions as nucleoside substitute in the synthesis of dimers and 13-mers. therefore we used the phosporamidite and the h-phosphonate chemistry. the nuclease susceptibilities and the base-pairing properties of these oligomers have been evaluated.	1991	2041735
analysis of acrylamido-buffers for isoelectric focusing by capillary zone electrophoresis.	immobilized ph gradients use a series of weak acrylamido acids and bases (immobiline) to create a ph gradient along the separation axis. these buffers can be degraded in water by two mechanisms: (i) hydrolysis of the amido bond, with generation of free acrylic acid and either an amino acid or a diamine; (ii) autopolymerization to oligomers and/or n-mers. in order to check for these degradation products, different capillary zone electrophoresis systems for analysis of all immobilines have been de ...	1991	2050100
[enzymatic ligation of dna fragments containing phosphoamide modification of the internucleotide bonds].	dna fragments with the point amidophosphate (cyclohexylamido- or morpholido-) modification in the sugar-phosphate backbone were synthesized and separated into individual diastereoisomer. the isomers were separated by the reversed-phase hplc (rpc), and chirality at phosphorus was assigned by a stereochemical correlation scheme using phosphorothioate standards. the rpc-retention time values for rp-isomers were found to be lower than for sp-analogues. amidophosphate dna fragments were used as p- an ...	1991	2064627
theoretical and experimental measures of dna helix stability and their relation to sequence specific repair of o6-ethylguanine lesions.	recent work (breslauer et al. (1986) proc. natl. acad. sci. (u.s.a.), 83, 3746) has provided a method for calculating empirical thermodynamic quantities for helix to coil transitions from the base sequence of any oligomer. it is shown in this work that the dna helix binding energy, calculated with the amber force field, for 9-mers of the type 5'-gggxgeyggg-3', where x and y are any base and the central ge is o6-ethylguanine, correlates well with the empirical delta g for helix to strand transiti ...	1991	2067552
large-scale economic synthesis of antisense phosphorothioate analogues of dna for preclinical investigations.	the therapeutic potential of antisense oligonucleotides will heavily depend on a balance of two factors: pharmacologic effectiveness and cost of production. pharmacologic optimization will be achieved to a limited degree in in vitro systems, but substantial progress can only be made in the context of appropriate in vivo models. the quantities of synthetic oligonucleotides required for modest in vivo testing are several thousandfold greater than can be produced by conventional dna synthesis techn ...	1990	2078018
[synthesis of oligoribonucleotides by the n-phosphonate method using alkali-labile 2'-o-protective groups. ii. various aspects of using 2'-o-benzoyl and anisole protective groups].	the n-acyl, 5'-o-trityl (meotr, (meo)2tr, me3tr), 2'-o-benzoyl (and anisole) nucleosides were prepared by selective aroylation of n,5'-protected nucleosides. by means of the reverse-phase microcolumn liquid chromatography it was shown that the rate of the aryl 2'----3'-isomerisation is lower in case of 2'-anisoylnucleosides and depends on structure of the 5'-o-protecting group. the prepared synthons were used for the manual h-phosphonate solid-phase synthesis of oligoribonucleotides (6-10-mers).	1990	2096827
oestrogen administration and the expression of the kallikrein gene family in the rat submandibular gland.	using a series of oligonucleotide probes (18-21 mers) specific for members of the rat kallikrein/tonin (arginyl-esteropeptidase) gene family (ps, s1, s2, s3, k1, p1), we have shown by northern blot analysis that all six genes are expressed in the submandibular gland (smg), with ps (true kallikrein) the most abundant in both male and female rats. though female levels of ps mrna are similar to that in the male, levels of mrna from both the kallikrein-like (s1, k1, p1) and tonin (s2)/tonin-like (s3 ...	1990	2155348
antisense oligodeoxyribonucleotides inhibit the expression of the gene for hepatitis b virus surface antigen.	the effect of a series of antisense oligodeoxyribonucleotide [oligo(dn)] on the expression of the surface antigen (hbsag) gene of human hepatitis b virus (hbv) was examined using hepatocellular carcinoma cells that contain integrated hbv genomes. of a number of antisense oligo(dn)s tested, synthetic 15-mers directed at the cap site of mrna and regions of the translational initiation site of the hbsag gene were found to be highly effective and inhibited viral gene expression by as much as 96%. th ...	1990	2177093
analysis of spontaneous and psoralen-induced salmonella typhimurium hisg46 revertants by oligodeoxyribonucleotide colony hybridization: use of psoralens to cross-link probes to target sequences.	an improved dna colony-hybridization method for the rapid characterization of salmonella typhimurium hisg46 revertants is described. oligodeoxyribonucleotides (15-mers) complementary to each of 6 possible transition or transversion mutations and an extragenic suppressor mutation, underlying the his+ phenotype, were prepared. optimal sequence discrimination was achieved by hybridizing 15-mers at the apparent dissociation temperature (td) for 2 h with chromosomal dna of revertant colonies affixed ...	1990	2179713
genetic reconstruction and characterization of the recombinant transacylase (e2b) component of bovine branched-chain alpha-keto acid dehydrogenase complex. implication of histidine 391 as an active site residue.	genetically altered transacylase (e2b) proteins of the bovine branched-chain alpha-keto acid dehydrogenase complex were overexpressed in escherichia coli and characterized. deletion by psti or bal31 digestion of the amino-terminal region of the inner-core domain (residues 175-421) beyond residue 209 resulted in a complete loss of transacylase activity. the enzyme assay was carried out using [1-14c]isovaleryl-coa and exogenous dihydrolipoamide as substrates. the removal of 4 residues (thr-ile-pro ...	1990	2198287
application of long synthetic oligonucleotides for gene analysis: effect of probe length and stringency conditions on hybridization specificity.	two different lengths of long unique synthetic oligonucleotide probes (37- and 48-mers) specific for human major histocompatibility complex (mhc) class ii beta genes were synthesized. these oligonucleotides were utilized to examine factors influencing hybridization specificity. both probe length and stringency of washing conditions were found to be crucial factors for sequence-specific hybridization.	1990	2206600
a novel protein binds a key origin sequence to block replication of an e. coli minichromosome.	a sequence of three tandem repeats of a 13-mer in the replication origin (oric) of e. coli is the highly conserved site of opening of the duplex for initiation of dna synthesis. a protein that binds this sequence has been discovered in e. coli and purified to homogeneity. this novel 33 kd polypeptide behaves as a dimer. binding to the 13-mers is specific and limited to this region. at a ratio of 10-20 monomers per oric plasmid, the binding blocks initiation by preventing the opening of the 13-me ...	1990	2208289
recombinant hnrnp protein a1 and its n-terminal domain show preferential affinity for oligodeoxynucleotides homologous to intron/exon acceptor sites.	the reported binding preference of human hnrnp protein a1 for the 3'-splice site of some introns (swanson and dreyfuss (1988) embo j. 7, 3519-3529; mayrand and pederson (1990) nucleic acids res. 18, 3307-3318) was tested by assaying in vitro the binding of purified recombinant a1 protein (expressed in bacteria) to synthetic oligodeoxynucleotides (21-mers) of suitable sequence. in such a minimal system we find preferential binding of protein a1 to oligodeoxynucleotide sequences corresponding to t ...	1990	2251120
detection and typing of maize streak virus and other distantly related geminiviruses of grasses by polymerase chain reaction amplification of a conserved viral sequence.	the application of the polymerase chain reaction dna amplification technique to the detection and typing of isolates of maize streak virus (msv) and other related geminiviruses of grasses is described. the oligonucleotide primers used for amplification were 17-mers which contained a number of degeneracies. an approximately 250 base pair fragment was amplified from all geminivirus-infected grass and cereal samples tested. the amplification reaction was specific, working down to a concentration of ...	1990	2254750
reca protein self-assembly. ii. analytical equilibrium ultracentrifugation studies of the entropy-driven self-association of reca.	we have investigated the self-association of reca protein from escherichia coli by equilibrium ultracentrifugation. monomeric reca (mr = 37,842) was observed in reversible equilibrium with trimers, hexamers and dodecamers in the presence of 1.5 m-kcl, 5 mm-hepes, 1 mm-edta, 2 mm-atp (ph 7.0) at 1 degrees c. the equilibrium was strongly temperature-dependent, with polymerization being favored as the temperature was raised from 1 degrees c 21 degrees c, and was reversible with respect to temperatu ...	1990	2266565
amplification of a long sequence that includes a processed pseudogene for elongation factor 2 in the mouse.	quantitative southern blotting analysis has demonstrated that mouse cells contain about 70 copies per haploid genome of a dna sequence related to the gene for elongation factor 2. the restriction maps of seven cosmids that each carry one copy of the ef2-related sequence (mer) and nucleotide sequences of mers were highly conserved among the cosmids. data obtained by such analyses suggest that mers were produced by the integration of one copy of mer derived from poly(a)+ mrna for ef2 into a specif ...	1990	2303263
sequence-dependent oligonucleotide-target duplex stabilities: rules from empirical studies with a set of twenty-mers.	we were interested in developing a better method to predict the thermal stability of specific oligonucleotide-target duplexes. recognizing that the base sequence can have important effects, we investigated the use of a simple parameter based on nearest-neighbor stacking interactions, the mean stacking temperature. we took values for doublet stabilities from the literature and used a computer program to calculate mean stacking temperatures for all oligonucleotides of specified length and g + c co ...	1990	2357384
thermal denaturation profiles and gel mobility shift analysis of oligodeoxynucleotide triplexes.	oligodeoxypurine- and oligodeoxypyrimidine-containing strands were mixed under conditions conducive to the formation of triple stranded assemblies. the mixtures were characterized both by their uv absorbance change with increasing temperature and by their mobility in non-denaturing polyacrylamide gels. duplexes 34 bp long containing 15 central purines on one strand and 15 complementary pyrimidines on the other strand yielded new melting transitions and showed different gel mobilities upon combin ...	1990	2395647
dynamics of dna polymerase iii holoenzyme of escherichia coli in replication of a multiprimed template.	movements of dna polymerase iii holoenzyme (holoenzyme) in replicating a template multiprimed with synthetic pentadecadeoxynucleotides (15-mers) annealed at known positions on a single-stranded circular or linear dna have been analyzed. after extension of one 15-mer on a multiprimed template, holoenzyme moves downstream in the direction of chain elongation to the next primer. holoenzyme readily traverses a duplex, even 400 base pairs long, to exploit its 3'-hydroxyl end as the next available pri ...	1985	2413035