Publications

sars hcov papain-like protease is a unique lys48 linkage-specific di-distributive deubiquitinating enzyme.	ubiquitin (ub) and the ub-like (ubl) modifier interferon-stimulated gene 15 (isg15) participate in the host defence of viral infections. viruses, including the severe acute respiratory syndrome human coronavirus (sars hcov), have co-opted ub-isg15 conjugation pathways for their own advantage or have evolved effector proteins to counter pro-inflammatory properties of ub-isg15-conjugated host proteins. in the present study, we compare substrate specificities of the papain-like protease (plpro) fro ...	0	25764917
middle east respiratory syndrome coronavirus: another zoonotic betacoronavirus causing sars-like disease.	the source of the severe acute respiratory syndrome (sars) epidemic was traced to wildlife market civets and ultimately to bats. subsequent hunting for novel coronaviruses (covs) led to the discovery of two additional human and over 40 animal covs, including the prototype lineage c betacoronaviruses, tylonycteris bat cov hku4 and pipistrellus bat cov hku5; these are phylogenetically closely related to the middle east respiratory syndrome (mers) cov, which has affected more than 1,000 patients wi ...	0	25810418
replication and shedding of mers-cov in upper respiratory tract of inoculated dromedary camels.	in 2012, a novel coronavirus associated with severe respiratory disease in humans emerged in the middle east. epidemiologic investigations identified dromedary camels as the likely source of zoonotic transmission of middle east respiratory syndrome coronavirus (mers-cov). here we provide experimental support for camels as a reservoir for mers-cov. we inoculated 3 adult camels with a human isolate of mers-cov and a transient, primarily upper respiratory tract infection developed in each of the 3 ...	0	25418529
the amino acids 736-761 of the mers-cov spike protein induce neutralizing antibodies: implications for the development of vaccines and antiviral agents.	based on a bioinformatics analysis of the middle east respiratory syndrome coronavvirus (mers-cov) s protein, we synthesized a panel of peptides coupled to keyhole limpet haemocyanin and used them to raise antibodies in rabbits. in addition, the recombinant receptor-binding domain (rbd) was used to raise polyclonal antibodies in mice. all of the antibodies raised by s-peptide immunisation were specific and sensitive for s protein expressed in transfected cells in the indirect immunofluorescence ...	0	25387086
induction and suppression of an autoimmune disease by oligomerized t cell epitopes: enhanced in vivo potency of encephalitogenic peptides.	t cell epitope peptides derived from proteolipid protein (plp139-151) or myelin basic protein (mbp86-100) induce experimental autoimmune encephalomyelitis (eae) in "susceptible" strains of mice (e.g., sjl/j). in this study, we show that the encephalitogenic effect of these epitopes when injected subcutaneously in complete freund's adjuvant was significantly enhanced if administered to the animal in a multimerized form as a t cell epitope oligomer (i.e., as multiple repeats of the peptide epitope ...	0	10684863
a synthetic consensus anti-spike protein dna vaccine induces protective immunity against middle east respiratory syndrome coronavirus in nonhuman primates.	first identified in 2012, middle east respiratory syndrome (mers) is caused by an emerging human coronavirus, which is distinct from the severe acute respiratory syndrome coronavirus (sars-cov), and represents a novel member of the lineage c betacoronoviruses. since its identification, mers coronavirus (mers-cov) has been linked to more than 1372 infections manifesting with severe morbidity and, often, mortality (about 495 deaths) in the arabian peninsula, europe, and, most recently, the united ...	0	26290414
replicative capacity of mers coronavirus in livestock cell lines.	replicative capacity of middle east respiratory syndrome coronavirus (mers-cov) was assessed in cell lines derived from livestock and peridomestic small mammals on the arabian peninsula. only cell lines originating from goats and camels showed efficient replication of mers-cov. these results provide direction in the search for the intermediate host of mers-cov.	0	24457147
control at hospital level of infections by methicillin-resistant staphylococci in children.	rapid spread of methicillin-resistant staphylococci (mers) in a children's hospital is described. within 4 months of the first isolation mers had been isolated from infections in all clinical units. mers were also regularly isolated at the out-patient department. protective isolation of one of the clinical units had no effect on the infection rate by mers. the use of antiseptics (hexachlorophene and chlorhexidine) and gentamicin nose cream in children and staff members in three out of five clini ...	1971	5285944
polyribosomes from peas: ii. polyribosome metabolism during normal and hormone-induced growth.	polyribosomes as large as 10-mers (strands of messenger rna bearing 10 ribosomes) were isolated from etiolated pea (pisum sativum l. var. alaska) stem tissue during all stages of development when methods were used which essentially eliminated ribonuclease activity during extraction. actively growing tissue, harvested from the apical 10 mm, yielded many large polyribosomes and a low (<20%) proportion of monosomes. similar tissue, allowed to age by applying lanolin to decapitated apices, showed a ...	1973	16658559
a motivation evaluating rating scale for chronic impaired schizophrenics (mers).	the attempt is made to structure a rating scale intended to evaluate the motivational level of chronic impaired schizophrenics (defect-schizophrenia). the need for such a measuring tool is stressed, considering mainly the relatively poor symptomatology of these patients, the difficulties to discern changes in the patients' condition and the fact that lack of motivation basically features the schizophrenic impairment. the scale strives to be as comprehensive as possible, extending from the lowest ...	1974	4469913
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
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
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
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
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
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
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
aggregation of small oligonucleosomal chains into 300-a globular particles.	chicken erythrocyte oligonucleosomes (trimers to about 20-mers) are able to interact with each other through the very lysine-rich histones (h1 and h5) and form heterogeneous globular particles with a mean diameter of about 300 a. these particles assemble spontaneously during micrococcal nuclease digestion of chromatin in the presence of 30 mm nacl and contain approximately 25 nucleosomes. they are sensitive to ionic strength and unfold at lower salt concentrations but can be reconstituted by res ...	1980	6935658
structural proteins of sarcophagid larval exoskeleton. composition and distribution of radioactivity derived from [7-14c]dopamine.	in the cyclorrhaphid flies, exoskeletal proteins from the last larval instar cross-link by arylation and glycosylation to form the sclerotized puparial case. cuticular proteins from maggots killed just prior to tanning were resolved into 21 soluble components by isoelectric focusing and sodium dodecyl sulfate-polyacrylamide electrophoresis. isoelectric points ranged from ph 4.5 to 6.0, molecular weights were distributed between mr = 16,000 and 24,000. aspartic and glutamic acids, glycine, serine ...	1981	7012149
hemocyanins in spiders, xv. the role of the individual subunits in the assembly of eurypelma hemocyanin.	the role of the seven different subunits in the quaternary structure of the 24-meric (37 s) hemocyanin of the tarantula, eurypelma californicum, was studied by reassembly experiments. individual subunits and combinations of 2, 3, 4, etc. different subunits were incubated in a total concentration of 1-2 mg/ml overnight in tris buffer, ph 7.5, omitting divalent cations. the reassembly mixtures were then analyzed by thinlayer gel filtration, electron microscopy, analytical ultracentrifugation, and ...	1982	7061045
cooperative binding of n-mers with steric hindrance to finite and infinite one-dimensional lattices.		1982	7082772
architecture of limulus polyphemus hemocyanin.	the architecture of the 48-meric hemocyanin of the horseshoe crab limulus polyphemus has been determined from electron micrographs of whole (48-mer) molecules and half- (24-mer) molecules. the assembly of hexamers of kidney-shaped subunits can produce two dodecameric enantiomorphs, designated as right and left. the assembly of 24-mers can again result in two enantiomorphs. by taking into account the rocking effect described by van heel and frank [van heel, m., & frank, j. (1981) ultramicroscopy ...	1982	7159564
isolation of intact polysomes from mechanically dehulled developing grain.	a rapid method for obtaining large quantities of developing groats suitable for the isolation of highly intact polysomes has been developed. developing spikelets were harvested directly from oat panicles into liquid nitrogen and then quickly passed through a dehuller. chaff was removed by air aspiration and the resultant groats were collected directly back into liquid nitrogen. approximately 250 g of groats could be isolated each man-hour by the above method. in comparison, only 10 g of endosper ...	1983	6650835
use of synthetic oligonucleotide probes complementary to genes for human hla-dr alpha and beta as extension primers for the isolation of 5'-specific genomic clones.	we have synthesized 175-nucleotide-long probes for the dna of human histocompatibility antigens hla-dr alpha and beta by extending on poly(a)+ mrna from b-cell lines with short synthetic deoxyribonucleotide primers complementary to the predicted nucleotide sequence of the nh2 terminus of both polypeptides. the synthesis of the probe for the alpha-chain dna was a two-step process starting with 11-mers which were extended by dideoxynucleotide chain termination experiments to a 20-mer of predicted ...	1983	6403940
isolation of polysomes from nostoc sp. mac and translation of messenger rna in a heterologous cell-free system.	a method has been established which isolated polysomes from the lysozyme/edta-shocked cyanobacterium, nostoc sp. mac. in a typical preparation the total recovery of rna as polysomes was 83%, in which 77% of the polysome fraction was present at sizes greater than 5-mers and 23% as 2-4-mers. messenger rna isolated from such a preparation of polysomes produced a 10-fold stimulation in the incorporation of [35s]methionine into polypeptides by a cell-free system of escherichia coli. the in vitro-synt ...	1983	6415228
size fractionation of thermal aggregates of immunoglobulin g.	purified pooled human immunoglobulin g (igg) in solution, when extensively heated at high temperatures or for long periods, irreversibly aggregates and insoluble precipitates result. however, when igg solutions are heated in the temperature range 55-65 degrees c for more limited time periods, soluble turbid polydispersed aggregate mixtures are obtained. gel filtration of such aggregate mixtures on calibrated bio-rad a-150m columns demonstrates a continuous size distribution from dimers to aggreg ...	1983	6660490
rapid synthesis of long-chain deoxyribooligonucleotides by the n-methylimidazolide phosphotriester method.	a modified phosphotriester method has been employed for the efficient chemical synthesis of long-chain deoxyribooligonucleotides. during the course of this work, a general and rapid procedure was developed for the preparation of 24-62-mers in solution. preparative reversed phase column chromatography on silanized silica gel was used to purify triester intermediates starting from 10-mers. the rapid synthesis of 32-mer and 42-mer on glass and silica gel supports using suitably protected 2-8-mer bl ...	1983	6324083
construction of improved m13 vectors using oligodeoxynucleotide-directed mutagenesis.	the restriction endonuclease cleavage sites for sphi and kpni have been added to the lac cloning region of the phage vectors m13mp10 and m13mp11, using oligodeoxynucleotide-directed in vitro mutagenesis. complementary deoxy 16-, 21- or 18-mers with the desired base changes were annealed to the m13mp dna strand and extended with the klenow fragment of dna polymerase i. in adding these sites we have shown that this technique can be used as a general method for inserting sequences of dna as well as ...	1983	6323249
simultaneous rapid chemical synthesis of over one hundred oligonucleotides on a microscale.	an inexpensive, extremely rapid manual method for simultaneous synthesis of large numbers of oligodeoxyribonucleotides on 50 or 150 nanomole scale is described. the oligonucleotides are assembled in parallel by the phosphotriester method on small cellulose paper disks in a simple gas pressure-controlled continuous-flow system. for each addition of a nucleotide the disks are sorted into four sets which are placed in four columns for addition of a, c, g and t, respectively. during one 2-week perio ...	1984	16453516
reactivity of sized thermal aggregates of immunoglobulin g with igm rheumatoid factor.	solutions of purified pooled human igg heated for varying times and temperatures between 55 degrees and 65 degrees yielded soluble turbid non-immune polydispersed aggregate mixtures. gel filtration of igg aggregate mixtures on calibrated biorad a-15, a-50 or a-150 columns resulted in continuous size distributions from dimers to particles with mol. wts. as high as 5 x 10(7) (300-mers) with no particular size predominant. chromatographically reproducible cuts of relatively narrow size heterogeneit ...	1984	6690393
immunological correspondence between arthropod hemocyanin subunits. i. scorpion (leiurus, androctonus) and spider (eurypelma, cupiennius) hemocyanin.	the hemocyanins of the scorpions leiurus quinquestriatus and androctonus australis, the tarantula eurypelma californicum (all 24-mers), and the lycosid spider cupiennius salei (dodecamer) were dissociated into subunits, the subunits isolated and studied by two-dimensional immunoelectrophoresis for interspecific cross-reactivities. androctonus hemocyanin yielded a pattern of 8 subunit types in agreement with data from lamy et al. (1979, arch. biochem. biophys. 193, 140-149). leiurus hemocyanin is ...	1984	6479892
genetic recombination of xenopus laevis 5 s dna in bacteria.	the behavior in genetic recombination of xenopus laevis 5 s dna has been examined, with particular emphasis on the role of 15-base-pair tandem repeats in the a + t-rich spacer. fragments of 5 s dna were introduced into escherichia coli cells as inserts in the recombination vectors, lambda rva and lambda rvb. intermolecular recombinants were selected in which, because of properties of the phage vectors, the crossover event must have occurred within the 5 s dna inserts. inserts from individual rec ...	1984	6092642
isolation and characterization of a cdna clone for the amino-terminal portion of the pro-alpha 1(ii) chain of cartilage collagen.	we have isolated a cdna clone (prcol 2) which is complementary to the 5'-terminal portion of the rat pro-alpha 1(ii) chain mrna. a synthetic oligonucleotide was used both as a primer for cdna synthesis and as a probe for screening a cdna library. the probe was a mixture of sixteen 14-mers deduced from an amino acid sequence present in the amino-terminal telopeptide of the rat cartilage alpha 1(ii) chain. this primer was chosen so that the resulting cdna would contain the sequence of the 5' end o ...	1984	6094525
use of complementary dna oligomers to probe trp leader transcript secondary structures involved in transcription pausing and termination.	dna oligomers were synthesized that are perfectly complementary to different segments of the tryptophan (trp) operon leader transcript. these 15 nucleotide long oligomers were used as probes of the involvement of transcript secondary structures in two processes: transcription pausing at the pause site located near base pair 90 in the leader region, and transcription termination at the attenuator. the 15-mers were complementary to the four segments of the trp leader transcript which have been sho ...	1984	6201827
substructure of human von willebrand factor.	using electron microscopy, we have visualized the substructure of human von willebrand factor (vwf) purified by two different approaches. vwf multimers, which appear as flexible strands varying in length up to 2 micron, consist of dimeric units (protomers) polymerized linearly in an end-to-end fashion through disulfide bonds. examination of small multimers (e.g., one-mers, two-mers, and three-mers) suggests that each protomer consists of two large globular end domains (22 x 6.5 nm) connected to ...	1985	2932468
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
[general approach to the engineering of synthetic dna].	a useful and efficient approach to the synthesis of dna duplexes of practically unlimited length has been developed. the proposed methodology is based on the use of temporary restriction sites for subcloning and assembling the segments of the desired dna. it allows the utilization of chemically synthesized oligonucleotides of various length (from 10- to 100-mers) for the duplex construction. the application of this approach to the synthesis of a gene for the functionally active bacteriorhodopsin ...	1985	3004509
in situ hybridization of putative somatostatin mrna within hypothalamus of the rat using synthetic oligonucleotide probes.	the distribution of mrna with high sequence homology to somatostatin mrna within the periventricular hypothalamus of rat was assessed using in situ hybridization techniques with synthetic oligodeoxyribonucleotide probes, complementary to the 3' coding region of rat somatostatin mrna. the probes (22- and 24-mers) were 5'-end labeled using t4 polynucleotide kinase and gamma-32p-atp. they were used either individually or after ligation with t4 dna ligase to form a 46-mer. serial tissue sections (le ...	1985	2860116
polyomavirus enhancer contains multiple redundant sequence elements that activate both dna replication and gene expression.	sequences that comprise the 244-base-pair polyomavirus enhancer region are also required in cis for viral dna replication (tyndall et al., nucleic acids res. 9:6231-6250, 1981). we have studied the relationship between the sequences that activate replication and those that enhance transcription in two ways. one approach, recently described by de villiers et al. (nature [london], 312:242-246, 1984), in which the polyomavirus enhancer region was replaced with other viral or cellular transcriptiona ...	1985	2985964
oligonucleotide probes for bacterial acylcarrier protein genes.	using a recently-introduced rapid manual method, we have synthesized a family of thirty six individual oligonucleotides of unique sequence (18-mers), which correspond to the conserved amino acid sequence, gadsld, found at the 4'-phosphopantetheine-binding site of the acylcarrier component of bacterial and plant fatty acid synthases. hybridisation of each of these oligonucleotides to southern blots of restricted streptomyces erythreus dna under stringent conditions showed that (i) only two probes ...	1985	4084606
site-specifically modified oligodeoxyribonucleotides as templates for escherichia coli dna polymerase i.	oligodeoxyribonucleotides with site-specific modifications have been used as substrates for escherichia coli dna polymerase i holoenzyme and klenow fragment. modifications included the bulky guanine-8-aminofluorene adduct and a guanine oxidation product resembling the product of photosensitized dna oxidation. by a combination of primers and "nick-mers", conditions of single-strand-directed dna synthesis and nick-translation could be created. our results show that the polymerase can bypass both t ...	1985	3887400
isolation of a cdna clone for the human lysosomal proteinase cathepsin b.	the cysteine proteinase cathepsin b is one member of the lysosomal acid hydrolases. based on the peptide sequence of rat liver cathepsin b, an oligonucleotide mixture containing 128 different 17-mers was synthesized and used as a probe to screen adult and fetal human liver cdna libraries. a recombinant clone with a 1540-nucleotide insert was identified from the fetal library, and dna sequence analysis confirmed that this clone encodes human cathepsin b. the clone, designated pcb-1, has sequences ...	1986	3010323
cc/gg contacts facilitate the b to a transition of dna in solution.	self-complementary decadeoxynucleotides, ccgatatcgg, ccagatctgg, ccctgcaggg, gggggccccc, were designed and synthesized to estimate the a-philic free energy of cc/gg contacts. first, regions of temperature-stability of the double-stranded conformation were determined for each 10-mer. then, circular dichroism spectra were recorded for the b-family forms at different temperatures, counter-ion concentrations and trifluoroethanol contents. a cooperative change typical of the b-a transition is observe ...	1986	2908426
xrep, a plasmid-stimulating x chromosomal sequence bearing similarities to the bk virus replication origin and viral enhancers.	the human x chromosome-linked fragment, "xrep," was sequenced because it exerts a positive effect on plasmid growth in both e. coli and saccharomyces cerevisiae. the sequence revealed three features similar to the human bk virus replication origin: xrep has a true palindrome, cctcc(t)3cctcc, which is similar to "true" palindrome-like sequences found at the replication origins of polyoma [cctc(t/c)10ctcc], bk [cctc(a/g)8cctcc] and sv40 [cctcc(a)6gcctcc] viruses. twenty nucleotides away from the t ...	1986	3025813
processivity and kinetics of the reaction of exonuclease i from escherichia coli with polydeoxyribonucleotides.	the enzyme exonuclease i from escherichia coli hydrolyzes successive nucleotides from the 3'-termini of single-stranded deoxyribonucleotide homopolymers. when the reaction is stopped after partial hydrolysis, only intact starting material and small oligomers can be isolated. the distribution of oligomeric products varies with the base composition of the polymer but the largest oligomer that can be isolated from the reaction of exonuclease i with homopolymers of deoxyadenylate, deoxythymidylate, ...	1986	3519606
two dr beta allelic series defined by exon ii-specific synthetic oligonucleotide genomic hybridization: a method of hla typing?	comparisons of exon ii hla-dr beta sequences have shown that nucleotide variations are principally clustered within the following three regions: v1 (amino acid 8-15), v2 (25-32), and v3 (70-77). v1, v2, and v3-derived 24-mers have been synthesized, the dr beta sequences coming from dr1, dr3, drw6, dr4, dr5, and drw53 haplotypes. each oligonucleotide was hybridized to pvu ii-digested dna samples from 13 hla genotyped families; therefore, 52 haplotypes have been investigated. six polymorphic pvu i ...	1986	3464000
use of oligonucleotide hybridization in the characterization of a beta zero-thalassemia gene (beta 37 tgg----tga) in a saudi arabian family.	analysis of restriction site polymorphisms in the beta-globin gene cluster of a saudi arabian female with beta zero-thalassemia demonstrated that both of her beta-globin genes were missing a nonpolymorphic avaii site in exon 2. examination of the normal nucleotide sequence surrounding this avaii site revealed that either of two nucleotide substitutions, tgg----tag or tgg----tga, could produce a nonsense codon at codon 37 and eliminate the avaii site. consequently, two oligonucleotides (19-mers s ...	1986	3006832
curved dna: design, synthesis, and circularization.	curved dna molecules and unusually small circles have been obtained by ligation of synthetic 21-base precursors: (sequence in text). the ligation resulted in the formation of double-stranded oligo-(precursor)s possessing a strong 10.5-base-pair (bp) periodicity of the runs of adenines. two-dimensional polyacrylamide gel electrophoresis of the ligation products showed two distinct families of spots: (i) noncircular oligo(precursor)s of 21 to 231 bp (1- to 11-mers) and (ii) four circles from 105 t ...	1986	3456570
binding of n-mers to one-dimensional lattices with longer than close-contact interactions.	a general formalism is derived for the evaluation of binding isotherms of n-mers (ligands) to one-dimensional polymers in the presence of ligand-ligand interactions which extend over several binding sites with distance-dependent interaction energies (multi-parameter model). this is an extension of the usual n-mer binding theory developed by several investigators in which ligand-ligand interaction occurs only when two ligands are in close contact (one-parameter model). the difference in binding i ...	1987	3607238
conformational transitions of synthetic dna sequences with inserted bases, related to the dodecamer d(cgcgaattcgcg).	conformational transitions for a series of imperfect palindromes related to the dodecamer d(cgcgaattcgcg) have been investigated. these sequences are: two isomeric 13-mers - d(cgcagaattcgcg) (13-meri) and d(cgcgaattacgcg) (13-merii), 17-mer d(cgcgcgaattacgcgcg) and 15-mer d(cgcgaaatttacgcg). insertion of a single adenine nucleotide prevents these sequences from being self-complementary. analysis of thermodynamic parameters derived from the melting profiles together with other data at higher conc ...	1987	3588311
an acridine-linked oligodeoxynucleotide targeted to the common 5' end of trypanosome mrnas kills cultured parasites.	anti-messenger oligodeoxynucleotides covalently linked to an intercalating agent were tested for their ability to inhibit translation of trypanosoma brucei mrnas in a cell-free system. the sequence of these oligodeoxynucleotides was complementary to part of the 35-nucleotide (nt) sequence which is present at the 5' end of all trypanosome mrnas (the so-called mini-exon sequence). in a rabbit reticulocyte lysate, a nonadeoxynucleotide linked to an acridine derivative, specifically inhibited protei ...	1987	3446576
a study of side reactions occurring during synthesis of oligodeoxynucleotides containing o6-alkyldeoxyguanosine residues at preselected sites.	as part of our studies on the molecular mechanisms of mutation by carcinogens we have synthesized 12 oligonucleotides (15-mers) containing an o6-alkylguanine residue at a preselected position for use as primers in the enzymatic synthesis of biologically active dna. ten of these oligonucleotides are derived from a minus strand sequence carrying the modified nucleotide in the third codon of gene g of bacteriophage phi x174 dna. two others are derived from plus strand sequences carrying the modific ...	1987	3607028
evidence that phosphorylation of eif-2(alpha) prevents the eif-2b-mediated dissociation of eif-2 x gdp from the 60 s subunit of complete initiation complexes.	recent observations have indicated that eukaryotic initiation factor (eif)-2 and gtp or gdp normally bind to 60 s ribosomal subunits in rabbit reticulocyte lysate and that when eif-2 alpha is phosphorylated and polypeptide chain initiation is inhibited, eif-2 x gdp accumulates on 60 s subunits due to impaired dissociation that is normally mediated by the reversing factor (eif-2b). current findings now indicate that inhibition due to phosphorylation of eif-2 alpha is mediated, at least in part, b ...	1987	3646234
comparison of the relative mutagenicities of o-alkylthymines site-specifically incorporated into phi x174 dna.	the relative mutagenicities of o-alkylthymine-dna adducts were analyzed in vivo by site-specific mutagenesis. purified dna polymerases were used to incorporate o4-methyl (me)-, o4-ethyl (et)-, o4-isopropyl (ipr)-, or o2-me-dttp onto the 3' terminus of a synthetic oligonucleotide (15-mer) hybridized to phi x174 am3 dna. the product oligonucleotides were further extended in the presence of unmodified dntps to yield 21-mers containing single o-alkylthymine adducts opposite the adenine residue of th ...	1987	2958453
antisense oligodeoxynucleotides provide insight into mechanism of translation initiation of two sendai virus mrnas.	translation of sendai virus nucleocapsid protein (np) and phosphoprotein (p/c) mrnas in rabbit reticulocyte lysates in the presence of antisense oligodeoxynucleotides (15-20-mers) showed that oligonucleotides having complementarity within the 5' noncoding region of the mrnas blocked translation, oligonucleotides having complementarity within 20 nucleotides upstream from the initiator codon blocked translation only partially, and oligonucleotides complementary to the coding region of mrna had no ...	1987	2438279
evaluation of n-ras oncogene anti-sense, sense and nonsense sequence methylphosphonate oligonucleotide analogues.	we have investigated the potential for using anti-sense non-ionic methylphosphonate oligonucleotide analogues to study the relationship between oncogene expression and maintenance of the transformed phenotype in malignant cells. our results confirmed that the methylphosphonates are resistant to biochemical degradation and are devoid of non-specific toxicity towards cultured human ht29 cells. at low temperature (less than 5 degrees c) both n-ras anti-sense and nonsense analogue 9-mers formed 1:1 ...	1988	2457379
the yeast uasg is a transcriptional enhancer in human hela cells in the presence of the gal4 trans-activator.	the yeast trans-activator protein gal4, when expressed in hela cells, stimulates transcription from several class b (ii) eukaryotic promoters containing gal4 binding sites either as the full uasg or as synthetic 17-mers. the characteristics of this activation are indistinguishable from those of the sv40 enhancer. transcription was similarly stimulated from either complex promoter regions containing multiple upstream elements or from a simple promoter region composed of only a tata box. addition ...	1988	2830022
comparative hybrid arrest by tandem antisense oligodeoxyribonucleotides or oligodeoxyribonucleoside methylphosphonates in a cell-free system.	antisense oligonucleotides containing either anionic diester or neutral methylphosphonate internucleoside linkages were prepared by automated synthesis, and were compared for their ability to arrest translation of human dihydrofolate reductase (dhfr) mrna in a nuclease treated rabbit reticulocyte lysate. in the case of oligodeoxyribonucleotides, tandem targeting of three 14-mers resulted in synergistic and complete selective inhibition of dhfr synthesis at a total oligomer concentration of 25 mi ...	1988	2836793
enzymatic amplification of myosin heavy-chain mrna sequences in vitro.	we have developed a procedure that detects the presence of mrna coding for human beta-myosin heavy chain in small amounts of total, unfractionated rna isolated from heart or skeletal muscle. the protocol is based on the enzymatic amplification in vitro of a selected 106-bp myosin isotype-specific subregion of this mrna. the method, which is a modification of the so-called "polymerase chain reaction," requires two synthetic oligonucleotide primers (20-mers), reverse transcriptase, and dna polymer ...	1988	2840250
a model for initiation at origins of dna replication.	many prokaryotic origins resemble e. coli oric in possessing essential at-rich sequences, tandemly repeated. the role of these repeats may be in the initial opening of the duplex by the initiator protein, as has been found for the 13-mers in oric and is implied for the 11-mers of the lambda origin. regulatory influences on the effective action of the initiator protein of e. coli (dnaa protein) include transcriptional activation of the origin, nucleotide binding and membrane attachment of the pro ...	1988	2843291
cytosine methylation does not affect binding of transcription factor sp1.	dna methylation may be a component of a multilevel control mechanism that regulates eukaryotic gene expression. we used synthetic oligonucleotides to investigate the effect of cytosine methylation on the binding of the transcription factor sp1 to its target sequence (a g + c-rich sequence known as a "gc box"). concatemers of double-stranded 14-mers containing a gc box successfully competed with the human metallothionein iia promoter for binding to sp1 in dnase i protection experiments. the prese ...	1988	3281160
optimal conditions for hybridization with oligonucleotides: a study with myc-oncogene dna probes.	we present a study on the refinement of filter-hybridization conditions for a series of synthetic oligonucleotides in the range from 17 to 50 base residues in length. experimental conditions for hybridization and the subsequent washing steps of the filter were optimized for different lengths of the synthetic oligonucleotides by varying the formamide concentration and washing conditions (temperature and monovalent cation concentration). target dna was immobilized to the nitrocellulose filter with ...	1988	3291638
functional mapping of the genome of the b19 (human) parvovirus by in vitro translation after negative hybrid selection.	we have analyzed the coding capacity of b19 parvovirus transcripts by in vitro translation using the negative hybrid selection technique. five different antisense oligonucleotides (18-mers) corresponding to different portions of the b19 genome were hybridized to rna samples extracted from human erythroid bone marrow cells infected with b19 parvovirus in vitro, and rnase h was added to cleave specific b19 rna molecules at selected sites. b19-specific translation products of these rna samples were ...	1988	3373576
[an improved chemico-enzymatic method of synthesizing long dna segments].	a simple and economy method of the biochemical assembling of long double-stranded dna segments is described. a single-stranded polydeoxynucleotide 122 bases long representing a fragment of synthetic gene of human beta-interferon was assembled from three synthetic fragments 36 (two) and 50 bases long on four complementary 12-mers as templates. this single-stranded polynucleotide was converted, in the presence of dna polymerase 1 and a 12-meric primer, in to the full-length double-stranded dna (th ...	1988	3382431
sequence-specific cleavage of rna using chimeric dna splints and rnase h.	to cleave rna molecules using e. coli rnase h in a site-specific manner, a short oligodeoxyribonucleotide (3-5 mer) linked with oligo(2'-o-methyl)ribonucleotide(s) was designed to be used as a dna splint. our model experiments with ribooligomer the splint duplexes (9 mers) and rnase h demonstrated that a tetradeoxynucleotide cluster seems to be sufficient for the enzyme recognition and the short dna-containing splint directs a unique cleavage of rna by rnase h. the method could be applied to lon ...	1988	2465538
cloning of macromomycin apoprotein gene from streptomyces macromomyceticus by use of 50-mer deoxynucleotide probes.	a mixed probe consisting of two synthetic deoxynucleotides (52 and 54 mers referred to as 50-mer) with arbitrarily chosen c or g for the third letters was prepared based on the amino acid sequences no. 31-48 and no. 72-90 of macromomycin (mcm) apoprotein and successfully used to clone the mcm apoprotein gene. digestion with sph i of total dna of mcm-producing streptomyces macromomyceticus m480-m1 yielded a 2.6-kb fragment that hybridized strongly to the probes. the hybridized probe was stable to ...	1988	3056895
sequence-specific binding and photocrosslinking of alpha and beta oligodeoxynucleotides to the major groove of dna via triple-helix formation.	a photocrosslinking reagent (p-azidophenacyl) was covalently linked to an octathymidylate synthesized with either the natural (beta) anomer of thymidine or the synthetic (alpha) anomer. the oligothymidylate was further substituted by an acridine derivative to stabilize the hybrid formed with a complementary octadeoxyadenylate sequence via intercalation. a single-stranded 27-mer containing a (da)8 sequence and a 27-mer duplex containing a (da.dt)8 sequence were used as targets. upon uv irradiatio ...	1988	3422738
construction of a series of several self-cleaving rna duplexes using synthetic 21-mers.	two fragments (21-mers) containing consensus sequences for the self-cleavage domain in transcripts of satellite dna of the newt were chemically synthesized and found to be cleaved in the presence of mg2+. the cleaved product contained the 3'-terminal 2',3'-cyclic phosphate. twenty-five combinations of partially double-stranded 21-mer rna which contained different bases within the consensus sequences and at the cleavage sites were tested for self-cleavage. it seemed that guanosine 3'-phosphate wa ...	1988	2449365
antisense oligonucleotide-directed cleavage of mrna in xenopus oocytes and eggs.	we have investigated the effect of specific antisense oligonucleotides on both exogenous and endogenous mrnas in xenopus oocytes and eggs. injection of 19- or 20-mers complementary to 70-kd heat shock protein, histone h4 and vegetally localized veg 1 coding sequences causes rapid cleavage and degradation of up to 96% of the target transcripts present in stage vi oocytes. nuclear and cytoplasmic transcripts appear to be equally accessible to cytoplasmically injected oligonucleotide and efficient ...	1988	2452730
synthesis of dna fragments containing 5,6-dihydrothymine, a major product of thymine gamma radiolysis.	5,6-dihydrothymine is one of the most important products of base damage by gamma irradiation of dna in anoxic conditions. this modified base is unstable in the deprotection conditions used for classical synthesis of oligonucleotides. for its incorporation in synthetic dna fragments, a new set of amino protecting groups has been developed. the 5,6-dihydrothymidine phosphoramidite was successfully employed for the synthesis of two 14-mers and one 17-mer bearing this defect at positions correspondi ...	1988	3340528
antisense oligodeoxyribonucleotide-directed cleavage of maternal mrna in xenopus oocytes and embryos.	we have investigated the effect of specific antisense oligodeoxynucleotides (oligos) on endogenous histone h4 mrna in xenopus oocytes, eggs and embryos. in unfertilised eggs and non-matured oocytes, one 20-mer oligo (h4-1) mediated the rnase h-like cleavage of up to 95% of h4 mrna (which included polysomal mrna), and cleavage was still obtained when the size of the oligo was reduced to a 10-mer; no cleavage was observed with 6- and 8-mers. the residual uncleaved mrna appeared to be completely in ...	1988	2468567
detection of nucleic acid hybridization by nonradiative fluorescence resonance energy transfer.	three approaches were used to study hybridization of complementary oligodeoxynucleotides by nonradiative fluorescence resonance energy transfer. (i) fluorescein (donor) and rhodamine (acceptor) were covalently attached to the 5' ends of complementary oligodeoxynucleotides of various lengths. upon hybridization of the complementary oligodeoxynucleotides, energy transfer was detected by both a decrease in fluorescein emission intensity and an enhancement in rhodamine emission intensity. in all cas ...	1988	3194390
[synthesis and cloning of artificial zein genes].	to investigate structure-function relationships in zein proteins and to develop an approach to the construction of mutant zeins of improved nutritional qualities, the chemical-enzymatic synthesis of a gene for zein cz22b1 (22 kd) has been undertaken. this 806-base pair long dna fragment consists of about 40 synthetic oligonucleotides, mostly 30-60-mers. the synthesis was planned with the use of a universal methodology for the artificial gene construction. the choice of appropriate sites for alte ...	1988	2853634
the b-z conformational transition in folded oligodeoxynucleotides: loop size and stability of z-hairpins.	the capacity to assume a left-handed conformation and the thermodynamics of loop formation in concentrated aqueous naclo4 have been investigated for the following palindromic sequences: d-(cgcgcgaaaaacgcgcg) (a5), d(cgcgcgtttttcgcgcg) (t5), d(cgcgcgtacgcgcg) (ta), and d(cgcgcgatcgcgcg) (at). the results show that (a) each oligomer assumes a z conformation upon exposure to increasing naclo4 concentrations; the salt concentration at the transition midpoint is 1.8 m for both a5 and t5 and 3 and 3.5 ...	1988	3219338
construction and analysis of monomobile dna junctions.	immobile dna junctions are complexes of oligomeric dna strands that interact to yield branched structures in which the branch point cannot migrate. this is achieved by minimizing the sequence symmetry in the flanking arms, so that base pairs lock at the branch site. here, we report the design, synthesis, and analysis of two semimobile junctions, structures in which a controlled extent of branch point migratory freedom is deliberately introduced. we have constructed two minimally symmetric four-a ...	1988	3191106
negative ion fast atom bombardment mass spectrometry of oligodeoxynucleotide carbamate analogs.	oligonucleotide analogs, in which carbamate rather than phosphodiester linkages form the backbone, have been analyzed by negative ion fast atom bombardment mass spectrometry. these oligodeoxynucleotides, dimers (982.4 daltons) to 11-mers (4356.5 daltons), show intense [m - h]- ions and some degree of cleavage of the sugar-carbamate backbone structure in repeating fashion on both sides of the carbonyl groups. sequencing of the shorter chain oligonucleotide analogs, up to six bases long, is demons ...	1988	3191249
interferon-induced early changes in nuclear protein interactions with the interferon consensus sequence.	a number of genes are transcriptionally regulated by interferons via a cis acting regulatory element (interferon consensus sequence). after incubation with hela cell nuclear extracts, a synthetic 29 mers probe representing the consensus was retarded as a single band whose amount increased after the addition of ifn to the cells. dnase i footprinting showed that a region of the non coding strand was protected differentially upon interferon treatment. after nuclear protein blotting and probing with ...	1989	2476126
structure-function relationships in a winter flounder antifreeze polypeptide. i. stabilization of an alpha-helical antifreeze polypeptide by charged-group and hydrophobic interactions.	the major antifreeze polypeptide (afp) from winter flounder (37 amino acid residues) is a single alpha-helix. aspartic acid and arginine are found, respectively, at the amino and carboxyl-termini. these charged amino acids are ideally located for stabilizing the alpha-helical conformation of this afp by means of charge-dipole interaction (shoemaker, k. r., kim, p.s., york, e.j., stewart, j. m., and baldwin, r. l. (1987) nature 326, 563-567). in order to understand these and other molecular inter ...	1989	2738067
allosteric oxygen-binding properties of reassembled tarantula (eurypelma californicum) hemocyanin with incorporated apo- or met-subunits.	4x6-meric hemocyanin of the tarantula eurypelma californicum was dissociated into subunits; one type of subunit was removed by immunoaffinity chromatography and replaced by its apo- or met-form. the mixture was reassembled and the reconstituted 4x6-mers were isolated. this was performed for subunits a, bc, d, e, f and g, respectively. it was verified by crossed immunoelectrophoresis that each type of subunit including the modified one, was incorporated in the reassembled 4x6-mers. oxygen binding ...	1989	2673295
site-specific interaction of intercalating drugs with a branched dna molecule.	the interaction of a stable branched dna molecule with an intercalative drug is probed by hydroxyl radical scission. methidiumpropyl-edta.fe(ii) [mpe.fe(ii)], consisting of an intercalating ring system tethered to edta.fe(ii), produces the hydroxyl radicals by means of a fenton reaction. the cleavage patterns of each labeled strand in a branched tetramer of four 16-mers are compared with those of the same strands in unbranched duplex controls. strong differences between the profiles correspondin ...	1989	2543439
effect of ionic strength on the hybridization of oligodeoxynucleotides with reduced charge due to methylphosphonate linkages to unmodified oligodeoxynucleotides containing the complementary sequence.	a 12-mer oligodeoxynucleotide containing 10 methylphosphonate bonds and 1 phosphodiester bond was shown to bind specifically to the restriction endonuclease fragment containing complementary dna in a southern blot. this 12-mer as well as 14-mer oligodeoxynucleotides containing 3 methylphosphonate and 10 phosphodiester bonds was used to examine the effect of reduced charge on the thermodynamics of binding to complementary dna or complementary oligodeoxynucleotides with additional nucleotides over ...	1989	2713356
use of edta derivatization to characterize interactions between oligodeoxyribonucleoside methylphosphonates and nucleic acids.	edta-derivatized oligonucleoside methylphosphonates were prepared and used to characterize hybridization between the oligomers and single-stranded dna or rna. the melting temperatures of duplexes formed between an oligodeoxyribonucleotide 35-mer and complementary methylphosphonate 12-mers were 4-12 degrees c higher than those of duplexes formed by oligodeoxyribonucleotide 12-mers as determined by spectrophotometric measurements. derivatization of the methylphosphonate oligomers with edta reduced ...	1989	2469462
synthesis, thermal stability and reactivity towards 9-aminoellipticine of double-stranded oligonucleotides containing a true abasic site.	a 13 mers abasic oligonucleotide was synthetized. it was therefore possible to compare thermal stability and reactivity of duplex oligonucleotides either with an apurinic/apyrimidinic site or without any lesion. an important decrease in the melting temperature appeared for duplexes with an abasic site. the chemical reaction of these modified oligonucleotides with the intercalating agent 9-aminoellipticine was studied by gel electrophoresis and by fluorescence. the formation of a schiff base betw ...	1989	2602153
an oligonucleotide hybridization approach to dna sequencing.	we have proposed a dna sequencing method based on hybridization of a dna fragment to be sequenced with the complete set of fixed-length oligonucleotides (e.g., 4(8) = 65,536 possible 8-mers) immobilized individually as dots of a 2-d matrix [(1989) dokl. akad. nauk sssr 303, 1508-1511]. it was shown that the list of hybridizing octanucleotides is sufficient for the computer-assisted reconstruction of the structures for 80% of random-sequence fragments up to 200 bases long, based on the analysis o ...	1989	2680594
[in vitro anti-hiv activity of phosphorothioate alpha-anomeric oligodeoxynucleotides].	oligonucleotide analogs consisting exclusively of alpha-anomeric deoxynucleoside units bridged with phosphorothioate linkages have been synthesized and tested in vitro as antiviral agents against human immunodeficiency virus (hiv) in human t cells. two 28-mers, an homopolymer alpha-s-dc28 and an oligomer alpha-s-anti-rev complementary to the initiation site of the regulatory viral gene rev exhibited antiviral activities comparable to those reported for the corresponding beta-anomeric phosphoroth ...	1989	2516764
sequencing of megabase plus dna by hybridization: theory of the method.	a mismatch-free hybridization of oligonucleotides containing from 11 to 20 monomers to unknown dna represents, in essence, a sequencing of a complementary target. realizing this, we have used probability calculations and, in part, computer simulations to estimate the types and numbers of oligonucleotides that would have to be synthesized in order to sequence a megabase plus segment of dna. we estimate that 95,000 specific mixes of 11-mers, mainly of the 5'(a,t,c,g)(a,t,c,g)n8(a,t,c,g)3' type, hy ...	1989	2737674
sequence analysis of rsk, a portion of the 95-kilobase plasmid of salmonella typhimurium associated with resistance to the bactericidal activity of serum.	increased sensitivity to killing by human serum complement occurs in salmonella typhimurium strains in which the 95-kilobase virulence plasmid is integrated into the chromosome. this phenotypic change appears to be due to alterations in plasmid gene expression and is reversed by the presence of an autonomous plasmid bearing a cloned region of the virulence plasmid. accordingly, this region has been termed rsk for reduced serum killing. sequence analysis of the region reveals that rsk is composed ...	1989	2645213
comparative inhibition of ras p21 protein synthesis with phosphorus-modified antisense oligonucleotides.	a rabbit reticulocyte lysate translation assay was used to quantitatively compare a series of antisense oligodeoxyribonucleotides (11-mers) having different internucleoside linkages and various degrees of complementarity (100-80%) with the start codon and downstream 8 bases of balb-ras p21 mrna. the oligomers had either contiguous phosphodiester, or alternating methylphosphonate-phosphodiester, or contiguous methylphosphonate, or contiguous phosphorothioate linkages. under the conditions used fo ...	1989	2679622
amplification of human papillomavirus dna sequences by using conserved primers.	the polymerase chain reaction has potential for use in the detection of small amounts of human papillomavirus (hpv) viral nucleic acids present in clinical specimens. however, new hpv types for which no probes exist would remain undetected by using type-specific primers for the polymerase chain reaction before hybridization. primers corresponding to highly conserved hpv sequences may be useful for detecting low amounts of known hpv dna as well as new hpv types. here we analyze a pair of primers ...	1989	2556429
chemical synthesis of the 24 rna fragments corresponding to hop stunt viroid.	a general and practical synthetic method of oligoribonucleotides (10-20 mers) by using the cyanoethyl phosphoramidite approach was described. in this experiment 9-phenylxanthen-9-yl (pix) and 9-(4-methoxy)phenylxanthen-9-yl (mox) groups were employed for the 5'-hydroxyls and tetrahydropyranyl (thp) group was used for the 2'-hydroxyl protecting groups. in addition, suitable acyl groups were introduced for the protection of the lactam functions of guanine and uracil moieties.	1989	2813059
the dnaa initiator protein binds separate domains in the replication origin of escherichia coli.	after binding to its four 9-mer boxes in the 245-base pair escherichia coli replication origin (oric), dnaa protein effects the formation of an "open complex" in an adjacent region made up of three 13-mers (bramhill, d., and kornberg, a. (1988) cell 52, 743-755). this open complex formation requires the atp form of dnaa protein assisted by hu protein (sekimizu, k., bramhill, d., and kornberg, a. (1987) cell 50, 259-265). we now provide direct evidence that dnaa protein binds the 13-mers, sequenc ...	1989	2539372
phosphorothioate oligodeoxynucleotides are potent sequence nonspecific inhibitors of de novo infection by hiv.	phosphorothioate homo-oligodeoxynucleotides have recently been found to protect ath-8 cells against the cytopathic effect of de novo infection by hiv. the effect is dose and chain-length dependent, with a maximum effect seen for 21-28-mers. we have now synthesized a series of phosphorothioate oligomers with mixed-based sequences and found that all of them have a dose-dependent cytoprotective effect that is maximal at an oligomer concentration of about 1-2 microm. the least effective sequences co ...	1989	2482061
the expression of the kallikrein gene family in the rat pituitary: oestrogen effects and the expression of an additional family member in the neurointermediate lobe.	abstract using a series of oligonucleotide probes (18 to 21 mers) specific for members of the rat kallikrein/tonin gene family (ps, s1, s2, s3, k1, p1), we have shown by northern blot analysis that the oestrogen-dependent kallikrein gene expressed in the male and female rat anterior pituitary is true kallikrein (ps). in addition, we have demonstrated that oestrogen treatment may also induce ps gene expression in the male and female rat neurointermediate lobe. none of the other five rat arginyl-e ...	1989	19210455
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
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
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
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
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
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