| magnetic circular dichroism study of cytochrome ba3 from thermus thermophilus: spectral contributions from cytochromes b and a3 and nanosecond spectroscopy of co photodissociation intermediates. | near-uv-vis magnetic and natural circular dichroism (mcd and cd) spectra of oxidized, reduced, and carbonmonoxy-complexed cytochrome ba3, a terminal oxidase from the bacterium thermus thermophilus, and nanosecond time-resolved mcd (trmcd) and cd (trcd) spectra of the unligated species formed after photodissociation of the co complex are presented. the spectral contributions of individual cytochromes b and a3 to the soret region mcd are identified. trmcd spectroscopy is used to follow the spin st ... | 1992 | 1327113 |
| stabilization of escherichia coli ribonuclease hi by strategic replacement of amino acid residues with those from the thermophilic counterpart. | thermus thermophilus ribonuclease h is exceptionally stable against thermal and guanidine hydrochloride denaturations as compared to escherichia coli ribonuclease hi (kanaya, s., and itaya, m. (1992) j. biol. chem. 267, 10184-10192). the identity in the amino acid sequences of these enzymes is 52%. as an initial step to elucidate the stabilization mechanism of the thermophilic rnase h, we examined whether certain regions in its amino acid sequence confer the thermostability. a variety of mutant ... | 1992 | 1328237 |
| optimization of long-distance pcr using a transposon-based model system. | the ability to amplify routinely long pcr products (5-25 kb) with high specificity and fidelity, regardless of target template sequence or structure, would provide significant benefits to genome mapping and sequencing endeavors. although occasional reports have described the generation of long pcr products, such results have been difficult to replicate and have frequently utilized probe hybridization to identify the specific product from nonspecific amplified dna. production of specific pcr prod ... | 1992 | 1337007 |
| cloning and sequencing of genes encoding the tthhb8i restriction and modification enzymes: comparison with the isoschizomeric taqi enzymes. | genes encoding the tthhb8i restriction and modification (r-m) system from thermus thermophilus hb8 (recognition sequence t decreases cga) were cloned in escherichia coli. the genes have the same transcriptional orientation, with the last 13 codons of the methyltransferase (mtase) overlapping the first 13 codons of the endonuclease (enase). nucleotide sequence analysis of the tthhb8i enase revealed a single chain of 263 amino acid (aa) residues that share a 77% identity with the corrected isoschi ... | 1992 | 1339363 |
| structure of holo-chaperonin studied with electron microscopy. oligomeric cpn10 on top of two layers of cpn60 rings with two stripes each. | a structural model of holo-chaperonin, known as a protein-folding control protein comprising 60 kda (cpn60) and 10 kda polypeptides (cpn10), is proposed based on the electron microscopic images of holo-chaperonin from thermus thermophilus and cpn60 from paracoccus denitrificans. isolated paracoccus cpn60 shows very similar images to those of escherichia coli tetradecameric cpn60, a seven-membered ring in the top view and a rectangular shape with four stripes in the side view. however, a small nu ... | 1992 | 1347504 |
| crystallization of the cpn60/cpn10 complex ('holo-chaperonin') from thermus thermophilus. | a stable complex of the chaperonins, cpn60 and cpn10 (escherichia coli groel and groes homologues), from the extremely thermophilic bacterium thermus thermophilus has been isolated and crystallized. the crystals have dimensions up to 30 x 200 x 200 microns. ultra-thin sections of the crystals estimated by electron microscopy showed a rectangular lattice with unit cell parameters of a = 17 nm, b = 27 nm, gamma = 90 degrees. | 1992 | 1356830 |
| highly sensitive and fast detection of phoma tracheiphila by polymerase chain reaction. | a new method for the diagnosis of the plant pathogenic fungus phoma tracheiphila has been developed. the method takes advantage of the enzymatic amplification of a specific 102 bp-long target sequence of fungal dna by the polymerase chain reaction (pcr) using thermus aquaticus dna polymerase. the amplified dna was characterized by agarose-gel electrophoresis, molecular hybridization using a synthetic oligonucleotide probe and direct sequencing. the application of the new method makes possible fa ... | 1990 | 1366441 |
| single chain antibody (sca) encoding genes: one-step construction and expression in eukaryotic cells. | we report the expression, in eukaryotic cells, of a gene encoding a single chain antibody (sca) and a rapid method for the construction of such genes. a sca directed against the aromatic dye fluorescein was synthesized from a gene constructed by means of the simultaneous use of four pcr primers and templates of both light and heavy chain immunoglobulin cdnas in the form of either plasmid clones or reverse transcribed hybridoma rna. two of the primers were partially complementary to one another a ... | 1991 | 1367186 |
| a universal method for the direct cloning of pcr amplified nucleic acid. | we have devised a simple, universal cloning strategy that permits the direct ligation of pcr amplified nucleic acid to a compatible vector preparation. the method does not require that special restriction sites or additional sequences be appended onto the amplification primers, nor the use of restriction endonucleases, modifying enzymes, or any purification procedures. this approach takes advantage of the single 3' deoxyadenylate extension that thermus aquaticus, thermus flavus, and thermococcus ... | 1991 | 1367662 |
| efficient production of thermostable thermus thermophilus xylose isomerase in escherichia coli and bacillus brevis. | the xylose (glucose) isomerase from the thermophile thermus thermophilus seems to have potential for the development of new isomerization processes using high temperatures and slightly acidic ph. the isomerase has an optimum temperature at 95 degrees c, and is also very stable at high temperatures. the optimum ph is around 7.0, close to where by-product formation is minimal. since thermus produces only a little of this useful isomerase, the production of the cloned gene in escherichia coli and b ... | 1992 | 1368014 |
| molecular cloning and nucleotide sequence of the aminopeptidase t gene of thermus aquaticus yt-1 and its high-level expression in escherichia coli. | aminopeptidase t (ap-t) is a metallo-dependent dimeric enzyme of thermus aquaticus yt-1, an extremely thermophilic bacterium. we cloned the ap-t gene from t. aquaticus yt-1 into escherichia coli using a synthetic oligonucleotide as a hybridization probe. the nucleotide sequence of the ap-t gene was found to encode 408 amino acid residues with gtg as a start codon. the molecular weight was calculated to be 44,820. the ap-t was overproduced in e. coli (about 5% of total soluble protein) when the s ... | 1990 | 1368580 |
| involvement of nh2-terminal pro-sequence in the production of active aqualysin i (a thermophilic serine protease) in escherichia coli. | aqualysin i is a heat-stable subtilisin-type protease produced by thermus aquaticus yt-1. the precursor of aqualysin i consists of four domains: an nh2-terminal signal peptide, an nh2-terminal pro-sequence, a protease domain, and a cooh-terminal pro-sequence. in escherichia coli cells harboring recombinant plasmid carrying the aqualysin i gene, proteolytic activity is obtained on treatment at 65 degrees c and mature enzyme is detected. in the case of mutant genes containing partial deletions in ... | 1991 | 1368764 |
| dna amplification fingerprinting of bacteria. | we have amplified short arbitrary stretches of total bacterial dna to produce highly characteristic and complex dna fingerprints. this dna amplification fingerprinting (daf) strategy involves enzymatic amplification of dna directed by a single arbitrary oligonucleotide primer. amplification produces a characteristic spectrum of products that is adequately resolved by polyacrylamide gel electrophoresis and visualized by silver staining. although daf is simple in concept, we found that amplificati ... | 1992 | 1369011 |
| purification and characterization of a thermostable carboxypeptidase (carboxypeptidase taq) from thermus aquaticus yt-1. | a thermostable carboxypeptidase, which we named carboxypeptidase taq, was purified from thermus aquaticus yt-1 and characterized. the molecular weight of the enzyme was estimated to be about 56,000 and 58,000 on sds-polyacrylamide gel electrophoresis and gel filtration, respectively, indicating that the enzyme has a monomeric structure. the optimum ph of the enzyme was 8.0, and the optimum temperature for the reaction was 80 degrees c. the enzyme activity was dependent on cobalt ion and was inhi ... | 1992 | 1369078 |
| the linear pcr reaction: a simple and robust method for sequencing amplified rrna genes. | the linear polymerase chain reaction was used to sequence amplified rna genes from strains of bacillus, thermus and legionella. the technique described is simple and reproducible and it works well with double standard product which has been peg precipitated directly from pcr reactions. | 1991 | 1370053 |
| electron image analysis of ribosomal subunits from thermus aquaticus. | electron micrographs of ribosomal subunits from the thermophilic bacterium thermus aquaticus were analysed using multivariate statistical analysis and characteristic views constructed to reproducible spatial resolutions ranging from 1.9 to 3.6 nm. these views were comparable to morphological classes of escherichia coli ribosomal subunits, albeit with differences in fine features also found in archaebacterial ribosomes. | 1992 | 1370377 |
| 2',5'-bis-o-(tert-butyldimethylsilyl)-3'-spiro-5''-(4''-amino-1'',2''- oxathiole-2'',2'-dioxide)pyrimidine (tsao) nucleoside analogues: highlyselective inhibitors of human immunodeficiency virus type 1 that are targeted at the viral reverse transcriptase. | a series of pyrimidine nucleoside analogues containing [2',5'-bis-o-(tert-butyldimethylsilyl)-3'-spiro-5''-(4''-amino- 1'',2''-oxathiole-2'',2''-dioxide)]-beta-d-ribofuranose as the pentose were found to inhibit human immunodeficiency virus type 1 [hiv-1(iiib)] replication at a concentration of 0.06-0.8 microm but were not cytotoxic at a 1000- to 10,000-fold higher concentration. these nucleoside derivatives were also effective against various other hiv-1 strains, including those resistant to 3' ... | 1992 | 1374900 |
| dna synthesis blocking lesions induced by singlet oxygen are targeted to deoxyguanosines. | in vitro dna synthesis on single stranded templates damaged by singlet oxygen was investigated in the supf trna gene sequence, using several dna polymerases. singlet oxygen was generated by the thermal decomposition of the water soluble with the endoperoxide of disodium 3,3'-(1,4-naphthylidene) dipropionate (ndpo2). the data demonstrated that damage at deoxyguanosine residues interrupts dna polymerization. modified t7 phage and thermus aquaticus dna polymerases were found to synthesize dna fragm ... | 1992 | 1375992 |
| determination of trna(phe) recognition nucleotides for phenylalanyl-trna synthetase from thermus thermophilus. | the trna(phe) recognition nucleotides for phenylalanyl-trna synthetase from an extreme thermophile thermus thermophilus were investigated. using yeast trna(phe) t7 transcripts with various point mutations it was shown that four recognition points (g34, a35, a36 from anticodon and a73 from acceptor stem) are important for aminoacylation at 37 degrees c. in the case of the 73rd discriminator base a----u, but not a----c, substitution suppresses aminoacylation. position 20, which proved to be essent ... | 1992 | 1379078 |
| 'run-off' polymerization with digoxigenin labelled nucleotides creates highly sensitive and strand specific dna hybridization probes: synthesis and application. | in this paper the in vitro synthesis and application of non-radioactively labelled strand specific dna probes is described. the probe is labelled by incorporation of nucleotides with the hapten digoxigenin into single-stranded dna during a 'run-off' reaction catalyzed by thermus aquaticus (taq) dna-polymerase. the 'run-off' reaction requires a linearized plasmid template and one primer binding site at a defined distance from the restriction site. single-stranded dna can be synthesized during rep ... | 1992 | 1381047 |
| a new crystal form of the complex between seryl-trna synthetase and trna(ser) from thermus thermophilus that diffracts to 2.8 a resolution. | two distinct complexes between seryl-trna synthetase and trna(ser) from thermus thermophilus have been crystallized using ammonium sulphate as a precipitant. form iii crystals grow from solutions containing a 1:2.5 stoichiometry of synthetase dimer to trna. they are of monoclinic space group c2 with unit cell dimensions a = 211.6 a, b = 126.8 a, c = 197.1 a, beta = 132.4 degrees and diffract to about 3.5 a. preliminary crystallographic results show that the crystallographic asymmetric unit conta ... | 1992 | 1397266 |
| cloning and sequence analysis of the phenylalanyl-trna synthetase genes (phest) from thermus thermophilus. | | 1992 | 1397273 |
| phenylalanyl-trna synthetase from thermus thermophilus can attach two molecules of phenylalanine to trna(phe). | phenylalanyl-trna synthetase from the extreme thermophilic bacterium thermus thermophilus can incorporate more than one molecule of phenylalanine into the trna(phe). it is shown that the 'hyperaminoacylated' trna(phe) is the bis-2',3'-o-phenylalanyl-trna(phe), and its formation is typical for the thermophilic enzyme but does not occur for e. coli phenylalanyl-trna synthetase under the same conditions. | 1992 | 1397314 |
| development of thermus-escherichia shuttle vectors and their use for expression of the clostridium thermocellum cela gene in thermus thermophilus. | we describe the self-selection of replication origins of undescribed cryptic plasmids from thermus aquaticus y-vii-51b (atcc 25105) and a thermus sp. strain (atcc 27737) by random insertion of a thermostable kanamycin adenyltransferase cartridge. once selected, these autonomous replication origins were cloned into the escherichia coli vector puc9 or puc19. the bifunctional plasmids were analyzed for their sizes, relationships, and properties as shuttle vectors for thermus-escherichia cloning. se ... | 1992 | 1400194 |
| crystallization and preliminary x-ray studies of a bacillus subtilis and thermus thermophilus hb8 chimeric 3-isopropylmalate dehydrogenase and thermostable mutants of it. | a new type of chimeric 3-isopropylmalate dehydrogenase (2t2m6t) was produced by expressing the fused gene of bacillus subtilis and thermus thermophilus. the enzyme shows heat stability intermediate between those of the parents. the crystal of the enzyme belongs to the space group of p3(2)21, with cell dimensions of a = b = 78.9 a and c = 158.9 a. two thermostable mutants of the chimeric enzyme were prepared by site-directed mutagenesis and then crystallized. | 1992 | 1400259 |
| extension of base mispairs by taq dna polymerase: implications for single nucleotide discrimination in pcr. | thermus aquaticus (taq) dna polymerase was used to measure the extension efficiency for all configurations of matched and mismatched base pairs at template-primer 3'-termini. the transition mispairs, a(primer).c, c.a, g.t, and t.g were extended 10(-3) to 10(-4)-fold less efficiently than their correctly paired counterparts. relative efficiencies for extending transversion mispairs were 10(-4) to 10(-5) for t.c and t.t, about 10(-6) for a.a, and less than 10(-6) for g.a, a.g, g.g and c.c. the tra ... | 1992 | 1408758 |
| [purification and characterization of alpha-glucosidase from an extreme thermophile, thermus thermophilus hb 8]. | a highly thermostable alpha-glucosidase (e c.3.2.1.20) from an extreme thermophile, thermus thermophilus hb 8, was purified to homogeneous by ammonium sulfate fractionation, deae-cellulose chromatography and preparative slab gel electrophoresis. the enzyme was purified 17 fold with 21% recovery of activity. the enzyme had a molecular weight of 67000 by sds-page. the isoelectric point was ph4.5 by ief on pag. the enzyme hydrolized p-nitrophenyl-alpha-glucoside (pn-pg), sucrose and maltose, but no ... | 1992 | 1413735 |
| molecular and functional properties of ef-tu from calderobacterium hydrogenophilum. | protein synthesis elongation factor tu (ef-tu) was purified from an extreme thermophilic hydrogen-oxidizing bacterium calderobacterium hydrogenophilum. the relative molecular mass of ef-tu. gdp was 51,000. the factor was heat stable and lost only 50% of its activity after heating at 80 degrees c for 5 min. native and reduced ef-tu or ef-tu. gdp contained one sh-reactive group. the elongation factors from c. hydrogenophilum and e. coli were shown to be immunologically identical. from the southern ... | 1992 | 1414127 |
| dna sequencing with thermus acquaticus dna polymerase and direct sequencing of polymerase chain reaction-amplified dna. 1988. | | 1992 | 1422071 |
| sequence of the s-layer gene of thermus thermophilus hb8 and functionality of its promoter in escherichia coli. | the nucleotide sequence of the slpa gene, which is responsible for the synthesis of the s-layer protein of thermus thermophilus hb8, is described. this gene is transcribed as a unit in which the coding region is preceded by a 127-base-long leader mrna sequence. the promoter region is also recognized by the rna polymerase of escherichia coli because of the presence of homologous -35 and -10 boxes. homologies with other promoters from thermus spp. are also presented. | 1992 | 1429468 |
| molecular characterisation of a dna ligase gene of the extremely thermophilic archaeon desulfurolobus ambivalens shows close phylogenetic relationship to eukaryotic ligases. | a 3382 bp fragment containing a gene for a dna ligase from the extremely thermophilic, acidophilic, and facultatively anaerobic archaeon (archaebacterium) desulfurolobus ambivalens was cloned and sequenced. the deduced amino acid sequence (600 amino acids, 67619 molecular weight) showed 30-34% sequence identity with the atp-dependent eucaryal (eukaryotic) dna ligases of schizosaccharomyces pombe, saccharomyces cerevisiae, the human dna ligase i, and with the vaccinia dna ligase. distant similari ... | 1992 | 1437556 |
| [the use of dna polymerase from thermus thermophilus]. | national dna polymerase from thermus thermophilus was used in polymerase chain reaction (pcr). the synthetic oligonucleotides (primers) to the basic structural hiv genes gag, env, the genome dna of donor peripheral blood lymphocytes were used, and the controls included the plasmid dna with cloned hiv genome and the genome dna of peripheral blood lymphocytes from hiv-infected persons confirmed by elisa and western blot analysis. the pcr technique and evaluation of the obtained results are describ ... | 1992 | 1441435 |
| identification of the gene encoding transcription factor nusg of thermus thermophilus. | the nusg gene of thermus thermophilus hb8 was cloned and sequenced. it is located 388 bp downstream from tufb, which is followed by the genes for ribosomal proteins l11 and l1. no equivalent to sece preceding nusg, as in escherichia coli, could be detected. the nusg gene product was overproduced in e. coli. a rabbit antiserum raised against the purified recombinant nusg reacted exclusively with one protein band of t. thermophilus crude extracts in western blot (immunoblot) analyses, and no cross ... | 1992 | 1447157 |
| cloning and sequence analysis of the phenylalanyl-trna synthetase genes (phest) from thermus thermophilus. | while crystals suitable for x-ray diffraction analyses are available of phenylalanyl-trna synthetase (phers) from the thermophilic bacterium thermus thermophilus, neither the primary structure of its constituent alpha and beta subunits nor the nucleotide sequence of the corresponding phes and phet genes were known. using specific oligonucleotides of conserved phes regions that were adapted to the t. thermophilus codon usage, we identified, cloned and subsequently sequenced the phest genes of thi ... | 1992 | 1451792 |
| molecular cloning and nucleotide sequence of the dna polymerase gene from thermus flavus. | | 1992 | 1454544 |
| some characteristics of a serine proteinase isolated from an extreme thermophile for use in kinetically controlled peptide bond synthesis. | | 1992 | 1476386 |
| discrimination of listeria monocytogenes from other listeria species by ligase chain reaction. | a ligase chain reaction assay based on a single-base-pair difference in the v9 region of the 16s rrna gene (16s rdna) was developed to distinguish between listeria monocytogenes and other listeria species. for this purpose, two pairs of primers were designed, with one primer of each pair being radioactively labeled. the ligated product was separated from the primers by denaturing polyacrylamide gel electrophoresis and then detected by autoradiography. to achieve a higher sensitivity, the 16s rdn ... | 1992 | 1482171 |
| purification and characterization of a thermostable proteinase isolated from thermus sp. strain rt41a. | thermus sp. strain rt41a produces an extracellular thermostable alkaline proteinase. the enzyme has a high isoelectric point (10.25-10.5) which can be exploited in purification by using cation-exchange chromatography. the proteinase was purified to homogeneity and has a molecular mass of 32.5 kda by sds/page. it is a glycoprotein, containing 0.7% carbohydrate as glucose equivalents, and has four half-cystine residues present as two disulphide bonds. maximum proteolytic activity was observed at p ... | 1992 | 1499549 |
| sequence of the tufa gene encoding elongation factor ef-tu from thermus aquaticus and overproduction of the protein in escherichia coli. | the sequence of the tufa gene from the extreme thermophilic eubacterium thermus aquaticus ep 00276 was determined. the gc content in third positions of codons is 89.5%, with an unusual predominance of guanosine (60.7%). the derived protein sequence differs from tufa- and tufb-encoded sequences for elongation factor tu (ef-tu) of thermus thermophilus hb8, another member of the genus thermus, in 10 of the 405 amino acid residues. three exchanges are located in the additional loop of ten amino acid ... | 1992 | 1499561 |
| [analogs of nucleotides, modified by a sugar residue and pyrimidine base, in a dna synthesis reaction, catalyzed by thermus aquaticus dna polymerase]. | substrate properties of dntp analogues in the dna synthesis reaction catalyzed by thermus aquaticus dna polymerase were studied. it was shown that most of dntp analogues which were known as terminators of dna synthesis of e. coli dna polymerase i were able to terminate dna synthesis catalyzed by thermus aquaticus dna polymerase. an interesting feature of thermus aquaticus dna polymerase was the ability to utilize 3'-azido-2',3'-dideoxythymidine triphosphate as terminating substrate. relative eff ... | 1992 | 1508169 |
| structure of the phenylalanyl-trna synthetase genes from thermus thermophilus hb8 and their expression in escherichia coli. | a 4459 bp long bamhi restriction fragment containing the two genes for the thermus thermophilus hb8 phenylalanyl-trna synthetase was cloned in escherichia coli and its nucleotide sequence was determined. the genes phes and phet encode the alpha- and beta-subunits with a molecular weight of 39 and 87 kd, respectively. three conserved sequence motifs typical for class ii trna synthetases occur in the alpha-subunit. secondary structure predictions indicate that an arm composed of two anti-parallel ... | 1992 | 1508711 |
| characterization of bacteriocins from two strains of bacillus thermoleovorans, a thermophilic hydrocarbon-utilizing species. | bacillus thermoleovorans s-ii and b. thermoleovorans nr-9 produce bacteriocins, and these bacteriocins are designated thermoleovorin-s2 and thermoleovorin-n9, respectively. the bacteriocins are effective against all but the producing strain of b. thermoleovorans, as well as being effective against salmonella typhimurium, branhamella catarrhalis, streptococcus faecalis, and thermus aquaticus. thermoleovorins are produced during log-phase growth and are inhibitory to actively growing cells. the ba ... | 1992 | 1514786 |
| three-dimensional structure of phenylalanyl-transfer rna synthetase from thermus thermophilus hb8 at 0.6-nm resolution. | the three-dimensional structure of the heterodimeric alpha 2 beta 2 enzyme phenylalanyl-trna synthetase from thermus thermophilus hb8 has been determined by x-ray crystallography, using the multiple-isomorphous-replacement method at 0.6 nm resolution. trigonal crystals of space group p3(2)21 have cell dimensions a = b = 17.6 nm and c = 14.2 nm. assuming one heterodimeric molecule/asymmetric unit, the ratio of unit cell volume/molecular mass was v = 0.00244 nm3/da, which is in the middle of the r ... | 1992 | 1521534 |
| molecular cloning of the isocitrate dehydrogenase gene of an extreme thermophile, thermus thermophilus hb8. | the gene coding for isocitrate dehydrogenase of an extreme thermophile, thermus thermophilus hb8, was cloned and sequenced. this gene consists of a single open reading frame of 1,485 bp preceded by a shine-dalgarno ribosome binding site. promoter- and terminatorlike sequences were detected upstream and downstream of the open reading frame, respectively. the g + c content of the coding region was 65.6%, and that of the third nucleotide of the codons was 90.3%. on the basis of the deduced amino ac ... | 1992 | 1539996 |
| glutamyl-trna synthetase from thermus thermophilus hb8. molecular cloning of the gltx gene and crystallization of the overproduced protein. | the gene for the glu-trna synthetase from an extreme thermophile, thermus thermophilus hb8, was isolated using a synthetic oligonucleotide probe coding for the n-terminal amino acid sequence of glu-trna synthetase. nucleotide-sequence analysis revealed an open reading frame coding for a protein composed of 468 amino acid residues (mr 53,901). codon usage in the t. thermophilus glu-trna synthetase gene was in fact similar to the characteristic usages in the genes for proteins from bacteria of gen ... | 1992 | 1541262 |
| structural analysis of three prokaryotic 5s rrna species and selected 5s rrna--ribosomal-protein complexes by means of pb(ii)-induced hydrolysis. | lead ions have been applied to the structural analysis of 5s rrna from thermus thermophilus, bacillus stearothermophilus and escherichia coli. based on the distribution of pb(ii)-induced cleavages, some minor modifications of the consensus secondary structure model of 5s rrna are proposed. they include the possible base pairing between nucleotides at positions 11 and 109, as well as changes in secondary interactions within the helix b region. the 'prokaryotic arm' region is completely resistant ... | 1992 | 1541273 |
| a polymerase chain reaction (pcr) method for sex and species determination with novel controls for deoxyribonucleic acid (dna) template length. | human x and y chromosome alpha-satellite sequences lying within higher order repeats were amplified by the polymerase chain reaction (pcr) in genomic deoxyribonucleic acid (dna) isolated from blood, bone, and several other tissues and specimens of potential forensic science interest. x and y sequences could be coamplified under some of the pcr conditions employed. monomorphic sequences in the 3'-apolipoprotein b gene (designated "h") and in an alpha-satellite higher order repeat on chromosome 17 ... | 1992 | 1545213 |
| streptococcus pneumoniae dna polymerase i lacks 3'-to-5' exonuclease activity: localization of the 5'-to-3' exonucleolytic domain. | the streptococcus pneumoniae pola gene was altered at various positions by deletions and insertions. the polypeptides encoded by these mutant pola genes were identified in s. pneumoniae. three of them were enzymatically active. one was a fused protein containing the first 11 amino acid residues of gene 10 from coliphage t7 and the carboxyl-terminal two-thirds of pneumococcal dna polymerase i; it possessed only polymerase activity. the other two enzymatically active proteins, which contained 620 ... | 1992 | 1548239 |
| nucleotide binding and gtp hydrolysis by elongation factor tu from thermus thermophilus as monitored by proton nmr. | proton nmr experiments of the gtp/gdp-binding protein ef-tu from the extremely thermophilic bacterium thermus thermophilus hb8 in h2o have been performed paying special attention to the resonances in the downfield region (below 10 ppm). most of these downfield signals are due to hydrogen bonds formed between the protein and the bound nucleotide. however, three downfield resonances appear even in the nucleotide-free ef-tu. the middle and c-terminal domain (domain ii/iii) of ef-tu lacking the gtp/ ... | 1992 | 1550822 |
| nmr study of the phosphate-binding loops of thermus thermophilus elongation factor tu. | the phosphoryl-binding loops in the guanosine diphosphate binding domain of elongation factor tu were studied by 15n heteronuclear proton-observe nmr methods. five proton resonances were found below 10.5 ppm. one of these was assigned to the amide group of lys 24, which is a conserved residue in the phosphoryl-binding concensus loop of purine nucleotide binding proteins. the uncharacteristic downfield proton shift is attributed to a strong hydrogen bond with a phosphate oxygen. the amide protons ... | 1992 | 1550823 |
| reasons and risks to study restriction/modification enzymes form extreme thermophiles: chilly coldrooms, 13th sample, and 13-codon overlap. | | 1992 | 1551586 |
| correlation between insertion mutant activities and amino acid sequence identities of the taqi and tthhb8 restriction endonucleases. | a two-codon insertion mutagenesis method has been generalized. over two dozen insertion mutants throughout the gene encoding taqi restriction endonuclease were constructed and activity was characterized. all mutants with activity either cleaved or nicked the canonical t decreases cga recognition sequence. some insertion mutants created duplication of gene regions, termed gemini proteins, which still retained activity. the correlation between mutants with poor activity and the regions of shared a ... | 1992 | 1551592 |
| the fidelity of taq polymerase catalyzing pcr is improved by an n-terminal deletion. | klentaq dna polymerase is an n-terminally truncated thermus aquaticus (taq) dna polymerase i. as expressed from a gene construct in escherichia coli, translation initiates at met236, bypassing the 5'----3' exonuclease domain of the dna polymerase-encoding gene. a sensitive forward mutation assay was used to measure the relative number of mutations introduced into the entire lacz gene by the polymerase chain reaction (pcr) under various conditions which allow the amplification of such a large dna ... | 1992 | 1551596 |
| the corrected nucleotide sequences of the taqi restriction and modification enzymes reveal a thirteen-codon overlap. | the nucleotide sequence of the genes encoding methyltransferase taqi (m.taqi) and restriction endonuclease taqi (r.taqi) with the recognition sequence, tcga, were analyzed in clones isolated from independent libraries. the genes, originally reported as 363 and 236 codons long [slatko et al., nucleic acids res. 15 (1987) 9781-9796] were redetermined as 421 and 263 codons long, respectively. the c terminus of the taqim gene overlaps the n terminus of the taqir gene by 13 codons, as observed with t ... | 1992 | 1551602 |
| crystallization of the seryl-trna synthetase-trna(ser) complex from thermus thermophilus. | the complex between seryl-trna synthetase and its cognate trna from the extreme thermophile thermus thermophilus has been crystallized from ammonium sulphate solutions. two different tetragonal crystal forms have been characterized, both diffracting to about 6 a using synchrotron radiation. one form grows as large bipyramids and has cell dimensions a = b = 127 a, c = 467 a, and the second form occurs as long, thin square prisms with cell dimensions a = b = 101 a, c = 471 a. analysis of washed an ... | 1992 | 1560467 |
| a rapid polymerase-chain-reaction-directed sequencing strategy using a thermostable dna polymerase from thermus flavus. | we have developed a polymerase chain reaction (pcr)-directed sequencing strategy for rapid sequencing of dna from crude viral or cell preparations. the basic strategy consists of two phases. in the first phase, the target dna is amplified by symmetric pcr with low concentrations of deoxyribonucleotide triphosphate (dntp) and oligodeoxyribonucleotide primers. this results in exponential amplification of dna in the initial cycles, reaching a plateau by 25 cycles due to limiting concentrations of d ... | 1992 | 1563631 |
| molecular cloning and nucleotide sequence of the gene encoding a h2o2-forming nadh oxidase from the extreme thermophilic thermus thermophilus hb8 and its expression in escherichia coli. | the sequence of the 32 n-terminal amino acids of the nadh oxidase from the extreme thermophile, thermus thermophilus hb8, was used to synthesize oligonucleotides to probe for the respective gene in a genomic library of t. thermophilus hb8. the gene encoding the nadh oxidase, designated nox, was cloned, its nucleotide sequence was determined and found to be colinear with the n-terminal sequence of the enzyme. the molecular mass of 26835 da, as deduced from the nox gene, agrees with that of the pu ... | 1992 | 1577004 |
| purification and characterization of a nadh oxidase from the thermophile thermus thermophilus hb8. | a nadh oxidase has been purified from the extreme thermophile thermus thermophilus hb8 by several chromatographic steps. the purified enzyme was essentially homogeneous as judged by gel electrophoresis under denaturing conditions and by determination of the n-terminal amino acids sequence. it is a monomeric flavin-adenine-dinucleotide-containing flavoprotein with an apparent molecular mass of 25 kda and an 1:1 ratio of fad to the polypeptide chain. the purified enzyme catalyzes the oxidation of ... | 1992 | 1577005 |
| n-acetylated and deacetylated 4'-fluoro-4-aminobiphenyl and 4-aminobiphenyl adducts differ in their ability to inhibit dna replication of single-stranded m13 in vitro and of single-stranded phi x174 in escherichia coli. | calf thymus single-stranded (ss) dna was modified with the n-sulfate conjugate of n-hydroxy-2-acetylaminofluorene (n-oh-aaf), n-hydroxy-4'-fluoro-4-acetylaminobiphenyl (n-oh-faabp) or n-hydroxy-4-acetylaminobiphenyl (n-oh-aabp) to yield predominantly n-acetylated adducts of 2-aminofluorene, 4-aminobiphenyl and 4'-fluoro-4-amino-biphenyl respectively to c8 of deoxyguanosine (dg-c8-aaf, dg-c8-aabp and dg-c8-faabp). the modified dnas were used as templates for in vitro dna synthesis. dna replicatio ... | 1992 | 1586987 |
| a comparative study of the relationship between thermostability and function of phenylalanyl-trna synthetases from escherichia coli and thermus thermophilus. | the relationship between thermostability and functional activities of phenylalanyl-trna synthetases (ec 6.1.1.20) from e. coli and thermus thermophilus has been studied. in the case of the e. coli enzyme, the activity decreased after the 43 degrees c treatment, both in the [32p]ppi-atp exchange reaction and the overall aminoacylation reaction, due to thermo-inactivation of the phenylalanyl-trna synthetase, whereas trna(phe) preserved its native structure. in the th. thermophilus system, the enzy ... | 1992 | 1587354 |
| overexpression of the thermus aquaticus b malate dehydrogenase-encoding gene in escherichia coli. | expression of the thermus aquaticus b malate dehydrogenase (mdh)-encoding gene (mdh), cloned in escherichia coli, was initially at a relatively low level (0.1% of soluble cell protein) and was effected by read-through from the tac promoter in the plasmid vector used. an enhancement in expression to 0.4% of soluble cell protein was achieved by shortening the intervening sequence between the promoter and the translation start codon of mdh. an ndei restriction site (5'-cat-atg-3') was engineered in ... | 1992 | 1587476 |
| affinity labeling of gtp-binding proteins in cellular extracts. | gtp-binding proteins in cellular extracts from escherichia coli, thermus thermophilus, yeast, wheat germ or calf thymus were identified using in situ periodate-oxidized [alpha-32p]gtp as affinity label. site-specific reaction of individual gtp-binding proteins was achieved by cross-linking the protein-bound 2',3'-dialdehyde derivative of gtp with the single lysine residue of the conserved nkxd sequence through schiff's base formation and subsequent cyanoborohydride reduction. labeled gtp-binding ... | 1992 | 1592117 |
| monofunctional dna-platinum(ii) adducts block frequently dna polymerases. | the question of whether monofunctional dna platinum(ii) adducts block synthesis of dna by purified dna polymerases of different types and origin has been investigated by comparing the time dependence of synthesis arrest and of dna adduct formation. activated salmon testis dna is used as a suitable substrate for dna synthesis allowing to probe inhibition by platinum(ii) monoadducts for the variety of inherent template-primers. reaction amplitudes are related to defined mixtures of dichloro and ch ... | 1992 | 1594449 |
| unusual resistance of peptidyl transferase to protein extraction procedures. | peptidyl transferase, the ribosomal activity responsible for catalysis of peptide bond formation, is resistant to vigorous procedures that are conventionally employed to remove proteins from protein-nucleic acid complexes. when the "fragment reaction" was used as a model assay for peptide bond formation, escherichia coli ribosomes or 50s subunits retained 20 to 40 percent activity after extensive treatment with proteinase k and sds, but lost activity after extraction with phenol or exposure to e ... | 1992 | 1604315 |
| the enzymes from extreme thermophiles: bacterial sources, thermostabilities and industrial relevance. | this review on enzymes from extreme thermophiles (optimum growth temperature greater than 65 degrees c) concentrates on their characteristics, especially thermostabilities, and their commercial applicability. the enzymes are considered in general terms first, with comments on denaturation, stabilization and industrial processes. discussion of the enzymes subsequently proceeds in order of their e.c. classification: oxidoreductases, transferases, hydrolases, lyases, isomerases and ligases. the ram ... | 1992 | 1605092 |
| conformational equilibrium of an enzyme catalytic site in the allosteric transition. | the dynamic equilibrium of a catalytic site between active and inactive conformations, the missing link between the structure and function of allosteric enzymes, was identified using protein engineering and nmr techniques. kinetic analyses of the wild-type and three mutants of thermus l-lactate dehydrogenase established that the allosteric property of the enzyme is associated with a concerted transition between the high-affinity (r) and low-affinity (t) states. by introducing mutations, we prepa ... | 1992 | 1606160 |
| rna catalysis. the universe expands. | | 1992 | 1608441 |
| a rapid method for preparation of bacterial chromosomal dna in agarose plugs using thermus rt41a proteinase. | | 1992 | 1616713 |
| a non-covalent nh2-terminal pro-region aids the production of active aqualysin i (a thermophilic protease) without the cooh-terminal pro-sequence in escherichia coli. | the precursor of aqualysin i, an extracellular protease produced by thermus aquaticus, consists of four domains: an n-terminal signal peptide, an n-terminal pro-sequence, the protease domain and a c-terminal pro-sequence. in an escherichia coli expression system, mature and active aqualysin i is formed by treatment at 65 degrees c and the n-pro-sequence is required for its production. complete deletion of the c-pro-sequence did not affect the production of active aqualysin i, indicating that the ... | 1992 | 1624114 |
| insertional mutagenesis in the extreme thermophilic eubacteria thermus thermophilus hb8. | the transcription and translation signals of the s-layer gene (slpa) from thermus thermophilus hb8 have been used to express a thermostable kanamycin adenyl transferase gene in this organism. the chimaeric resistance gene was inserted in vitro into slpa to produce different inactive forms of the gene, which were used to transform t. thermophilus hb8. after 48 hours of incubation at 70 degrees c, only two constructions that contained the kat gene flanked by thermus sequences from both sides of sl ... | 1992 | 1625584 |
| polymerase chain reaction amplification of single-stranded dna containing a base analog, 2-chloradenine. | by utilization of polymerase chain reaction techniques, single-stranded dna of defined length and sequence containing a purine analog, 2-chloroadenine, in place of adenine was synthesized. this was accomplished by a combination of standard polymerase chain amplification reactions with thermus aquaticus dna polymerase in the presence of four normal deoxynucleoside triphosphates, m13 duplex dna as template, and two primers to generate double-stranded dna 118 bases in length. an asymmetric polymera ... | 1992 | 1632514 |
| ribosomal proteins from thermus thermophilus for structural investigations. | in parallel with crystallographic studies of ribosomes from thermus thermophilus, a long-term program on the crystallization and structural investigations of ribosomal proteins from the same microorganism has been started at the institute of protein research (pushchino, russia). at present, more than half of the individual ribosomal proteins from t thermophilus have been purified without denaturating agents on a preparative scale and some of them have been obtained in the crystalline form. x-ray ... | 1992 | 1637860 |
| molecular cloning of a ribonuclease h (rnase hi) gene from an extreme thermophile thermus thermophilus hb8: a thermostable rnase h can functionally replace the escherichia coli enzyme in vivo. | a dna fragment encoding ribonuclease h (ec 3. 1.26.4) was isolated from an extreme thermophilic bacterium, thermus thermophilus hb8, by its ability to complement the temperature-sensitive growth of an escherichia coli rnha deficient mutant. the primary amino acid sequence showed 56% similarity to that of e. coli rnase hi but little or no homology to e. coli rnase hii. enzymes derived from thermophilic organisms tend to have fewer cysteines than their bacterial counterparts. however, t. thermophi ... | 1991 | 1653414 |
| the role of histidine-118 of inorganic pyrophosphatase from thermophilic bacterium ps-3. | treatment of the inorganic pyrophosphatase from thermophilic bacterium ps-3 with diethyl pyrocarbonate resulted in the almost complete loss of its activity, which followed pseudo-first-order kinetics. the presence of mg2+ prevented the inactivation. enzyme inactivated with diethyl pyrocarbonate was re-activated by hydroxylamine. the inactivation parallelled the amount of modified histidine residue, and a plot of the activity remaining against the amount of modified histidine residue suggested th ... | 1991 | 1654888 |
| magnetization of manganese superoxide dismutase from thermus thermophilus. | the ground state magnetic properties of manganese superoxide dismutase from thermus thermophilus in its native and reduced forms have been determined using saturation magnetization data. parallel epr measurements were used to verify that commonly encountered paramagnetic impurities were at low concentration relative to the metalloprotein. the native enzyme contains high spin mn(iii) (s = 2) with d = +2.44(5) cm-1 and e/d = 0. the reduced enzyme contains high spin mn(ii) (s = 5/2) with d = +0.50( ... | 1991 | 1655035 |
| a chaperonin from a thermophilic bacterium, thermus thermophilus, that controls refoldings of several thermophilic enzymes. | a chaperonin has been purified from a thermophilic bacterium, thermus thermophilus. it consists of two kinds of proteins with approximate mr 58,000 and 10,000 and shows a 7-fold rotational symmetry from the top view and a "football"-like shape from the side view under the electron microscopic view. its weak atpase activity is inhibited by sulfite and activated by bicarbonate. atp causes change of its mobility in nondenaturating polyacrylamide gel electrophoresis. the t. thermophilus chaperonin c ... | 1991 | 1682319 |
| image analysis by electron microscopy of two-dimensional crystals developed on a mercury surface of chaperonin from thermus thermophilus. | two-dimensional crystals of functional chaperonin molecules, which are protein complexes of cpn60 and cpn10, isolated from thermus thermophilus were prepared on a mercury surface under oxygen atmosphere and were observed by electron microscope after transferring them to carbon coated specimen grids. the crystals showed the hexagonal lattice with unit cell dimensions of a = b = 12.4 nm and gamma = 120 degrees. the averaged image at a 3 nm resolution of the chaperonin has a doughnut-like shape whi ... | 1991 | 1686608 |
| "run-off" synthesis and application of defined single-stranded dna hybridization probes. | a simple and efficient method for synthesizing radioactively labeled single-stranded dna hybridization probes with thermus aquaticus (taq) dna polymerase is described. this is done in a "run-off" polymerization with repeated cycles of denaturation, annealing, and extension. it leads to high yields of a single-stranded dna of defined length (up to 5000 nt), which is labeled to a high specific activity (1.3 x 10(8) cpm/micrograms dna). these hybridization probes are equally sensitive as nick-trans ... | 1990 | 1693048 |
| [the amplification of nucleotide sequences by pcr and the new technics for molecular diagnosis]. | the polymerase chain reaction (pcr) is a method for selective amplification of dna or rna segments of up to 2 kilobase pairs (kb) or more in length. repeated cycles of enzymatic primer extension in opposite directions are performed using synthetic oligonucleotides flanking the sequences of interest and the heat-stable dna polymerase from the archaebacterium thermus aquaticus (taq polymerase). the reaction is simple, fast and extremely sensitive, and the dna or rna content of a single cell is suf ... | 1990 | 1698384 |
| cdna library construction from small amounts of unfractionated rna: association of cdna synthesis with polymerase chain reaction amplification. | we describe here a general and simple procedure for cdna library construction making use of in vitro amplification of cdna by polymerase chain reaction (pcr). the first-strand cdna is synthesized from total rna with a primer ecori-(dt)17 and oligo(dg) tailed. an oligonucleotide, ecori-bamhi-(dc)13, is used to prime the second-strand synthesis by the thermostable dna polymerase of thermus aquaticus. the double-stranded cdna is then amplified directly by pcr. a study of the effect of the elongatio ... | 1990 | 1699456 |
| analysis and nucleotide sequence of the genes encoding the surface-layer glycoproteins of the hyperthermophilic methanogens methanothermus fervidus and methanothermus sociabilis. | the genes (slga) encoding the surface-layer glycoproteins of the hyperthermophilic methanogens methano-thermus pervidus and methanothermus sociabilis were cloned and sequenced. the nucleotide sequences of these genes differ at only nine positions, resulting in three amino acid differences. in both organisms, the transcription start site was localized by primer extension analyses. the dna sequence at this site conforms to the promotor box b motif for promotors of archaea. 24 nucleotides upstream ... | 1991 | 1712296 |
| reverse transcription and dna amplification by a thermus thermophilus dna polymerase. | a recombinant dna polymerase derived from the thermophilic eubacterium thermus thermophilus (tth pol) was found to possess very efficient reverse transcriptase (rt) activity in the presence of mncl2. many of the problems typically associated with the high degree of secondary structure present in rna are minimized by using a thermostable dna polymerase for reverse transcription, and predominantly full-length products can be obtained. the cdna can also be amplified in the polymerase chain reaction ... | 1991 | 1714296 |
| analysis of the gene encoding the rna subunit of ribonuclease p from t. thermophilus hb8. | the gene for the rna subunit of ribonuclease p from the extreme thermophilic eubacterium t. thermophilus hb8 was cloned using oligonucleotide probes complementary to conserved regions of rnase p rna subunits from proteobacteria. the monocistronic gene and its flanking regions were sequenced. the gene is enclosed by a promoter and a rho-independent terminator. nuclease s1 protection analyses showed that the primary transcript is identical with the mature rna, i.e. no processing events are involve ... | 1991 | 1719485 |
| thermus thermophilus ribosomes for crystallographic studies. | three-dimensional crystals of the 70s ribosomes, the 70s ribosome-mrna-trna complex, the 30s ribosomal subunits, several ribosomal proteins, the elongation factor g and threonyl- and seryl-trna synthetases from a gram-negative extreme thermophilic bacterium, thermus thermophilus, have been obtained at our institute. x-ray and neutronographic data from the 70s ribosome crystals have been collected up to 18 a and 60 a, respectively. two-dimensional crystalline sheets of the 70s ribosomes have been ... | 1991 | 1720668 |
| cell-free translation system using phosphorothioate-containing mrna. | phosphorothioate-containing rnas were generated by transcription of template dna using the sp diastereomers of ribonucleoside 5'-o-(1-thiotriphosphates) (ntp alpha s) and t7 rna polymerase. the substitution of mrna by phosphorothioate increased the efficiency of protein synthesis by stabilizing the mrnas in prokaryotic cell-free translation systems. the substituted mrnas were also shown to be applicable to the continuous cell-free translation system developed by spirin and coworkers. | 1991 | 1726804 |
| effects of different dna polymerases in ligation-mediated pcr: enhanced genomic sequencing and in vivo footprinting. | we have developed a simplified procedure for the ligation-mediated polymerase chain reaction (lmpcr) using thermococcus litoralis dna polymerase (vent dna polymerase). we show that vent dna polymerase produces correct, blunt-ended primer extension products with substantially higher efficiency than thermus aquaticus (taq) dna polymerase or modified t7 dna polymerase (sequenase). this difference leads to significantly improved genomic sequencing, methylation analysis, and in vivo footprinting with ... | 1992 | 1736283 |
| sequence and base modifications of two phenylalanine-trnas from thermus thermophilus hb8. | | 1992 | 1738592 |
| overproduction of the thermus thermophilus elongation factor tu in escherichia coli. | the elongation factor tu (ef-tu) encoded by the tufl gene of the extreme thermophilic bacterium thermus thermophilus hb8 was expressed under control of the tac promoter from the recombinant plasmid peftu-10 in escherichia coli. thermophilic ef-tu-gdp, which amounts to as much as 35% of the cellular protein content, was separated from the e coli ef-tu-gdp by thermal denaturation at 60 degrees c. the overproduced e coli-born t thermophilus ef-tu was characterized by: i) recognition through t therm ... | 1991 | 1742348 |
| the interplay between x-ray crystallography, neutron diffraction, image reconstruction, organo-metallic chemistry and biochemistry in structural studies of ribosomes. | crystals of ribosomes, their complexes with components of protein biosynthesis, their natural, mutated and modified subunits, have been subjected to x-ray and neutron crystallographic analyses. electron microscopy and 3-dimensional image reconstruction, supported by biochemistry, genetic, functional and organo-metallic studies were employed for facilitating phasing of the crystallographic data. for example, a monofunctional multi heavy-atom cluster (undecagold) was designed for covalent and quan ... | 1991 | 1742363 |
| analysis of mutations using pcr and denaturing gradient gel electrophoresis. | denaturing gradient gel electrophoresis (dgge) separates dna molecules based on primary sequence. under the appropriate conditions, all base pair (bp) substitutions, frame-shifts, and deletions less than about 10 bp can be resolved from the wild type sequence using dgge. polymerase chain reaction (pcr) permits facile amplification of a given region of the genome. we have combined pcr and dgge to: (i) localize mutations in the x-linked human androgen receptor gene. pcr/dgge was used to screen the ... | 1991 | 1748086 |
| three-dimensional structure of a highly thermostable enzyme, 3-isopropylmalate dehydrogenase of thermus thermophilus at 2.2 a resolution. | the three-dimensional structure of the highly thermostable 3-isopropylmalate dehydrogenase (ipmdh) from thermus thermophilus has been determined by the multiple isomorphous replacement method and refined to 2.2 a resolution. the final r-factor is 0.185 for 20,307 reflections. the crystal asymmetric unit has one subunit consisting of 345 amino acid residues. the polypeptide chain of this subunit is folded into two domains (first and second domains) with parallel alpha/beta motifs. the domains are ... | 1991 | 1748999 |
| [elimination of introns from the interleukin 1 beta genome by dna amplification and expression of interleukin 1 beta in escherichia coli]. | the use of the polymerase chain reaction was proposed for intron excision from genomic genes with known nucleotide sequences. three exons (5, 6 and 7) of genomic interleukin 1 beta gene were amplified by means of thermostable dna polymerase tthi from thermus thermophilus on the base of cloned in m13 phage human genomic interleukin 1 beta gene. synthetic oligonucleotides complementary to sequences flanking exons were used as primers. the fragments obtained by exon dna amplification were joined in ... | 1991 | 1753956 |
| cloning, overexpression and nucleotide sequence of a thermostable dna ligase-encoding gene. | thermostable dna ligase has been harnessed for the detection of single-base genetic diseases using the ligase chain reaction [barany, proc. natl. acad. sci. usa 88 (1991) 189-193]. the thermus thermophilus (tth) dna ligase-encoding gene (ligt) was cloned in escherichia coli by genetic complementation of a ligts 7 defect in an e. coli host. nucleotide sequence analysis of the gene revealed a single chain of 676 amino acid residues with 47% identity to the e. coli ligase. under phoa promoter contr ... | 1991 | 1756968 |
| sequence and expression of the gene encoding 3-phosphoglycerate kinase from bacillus stearothermophilus. | the structural gene (pgk) encoding 3-phosphoglycerate (pgk) from bacillus stearothermophilus nca1503, has been cloned in escherichia coli and its complete nucleotide sequence determined. the gene consists of an open reading frame corresponding to a protein of 394 amino acids (aa) (calculated mr 42,703) and, in common with other prokaryotic pgk genes, is preceded by the structural gene encoding glyceraldehyde-3-phosphate dehydrogenase (gapdh). constructs containing the b. stearothermophilus pgk g ... | 1991 | 1756980 |
| comparative analysis of ribosomal protein l5 sequences from bacteria of the genus thermus. | the genes for the ribosomal 5s rrna binding protein l5 have been cloned from three extremely thermophilic eubacteria, thermus flavus, thermus thermophilus hb8 and thermus aquaticus (jahn et al, submitted). genes for protein l5 from the three thermus strains display 95% g/c in third positions of codons. amino acid sequences deduced from the dna sequence were shown to be identical for t flavus and t thermophilus, although the corresponding dna sequences differed by two t to c transitions in the t ... | 1991 | 1764514 |
| [studies on the thermostable l-lactate dehydrogenase from thermophilic bacteria]. | about 200 strains of extreme thermophilic bacteria were isolated from hot springs in guandong province. a strain, hg25, was found to produce thermostable intracellular l-lactate dehydrogenase (ec. 1.1.1.27). it has the characteristic of thermus sp. the cells were gram-negative, non-sporulating, nonmotile, aerobic rods containing yellow pigment. the optimum temperature for growth was between 65 degrees c to 75 degrees c, the maximum 85 degrees c, and minimum 40 degrees c. the generation time at t ... | 1991 | 1814047 |
| molecular cloning of phosphofructokinase 1 gene from a thermophilic bacterium, thermus thermophilus. | phosphofructokinase 1 (pfk1) from thermus thermophilus differs from other bacterial pfks in that it is regulated by effector-induced reversible tetramer-dimer conversion rather than conformational change alone. we have cloned its gene and the deduced amino acid sequence was compared with that of other pfks. while almost all amino acid residues involved in substrate binding sites, which are assigned from crystal structures of other pfks, are well conserved, some possibly important changes are fou ... | 1991 | 1828151 |
| phosphoenolpyruvate-insensitive phosphofructokinase isozyme from thermus thermophilus hb8. | in the previous paper [xu, j., oshima, t., & yoshida, m. (1990) j. mol. biol. 215, 597-606], we reported that phosphofructokinase from thermus thermophilus is allosterically inhibited by phosphoenolpyruvate, which induces dissociation of the active four-subunit enzyme into an inactive two-subunit form. when t. thermophilus was cultured in a glucose-containing medium, another phosphofructokinase (pfk2) appeared in addition to the reported one (pfk1). the molecular weights of the native pfk2 molec ... | 1991 | 1830879 |
| molecular cloning of genes encoding major two subunits of a eubacterial v-type atpase from thermus thermophilus. | the atpab genes which encode the alpha and beta subunits of membrane atpase from a thermophilic eubacterium, thermus thermophilus hb8, were cloned. the deduced amino-acid sequences of the alpha subunit (583 amino acids) and the beta subunit (478 amino acids) are only moderately similar to the alpha beta subunits of the f0f1-atpases, while they are highly similar to the major two subunits of the v-type atpases, a family of atpases which have been so far found in eukaryotic endomembrane vacuolar v ... | 1991 | 1836357 |
| cloning, nucleotide sequence, and engineered expression of thermus thermophilus dna ligase, a homolog of escherichia coli dna ligase. | we have cloned and sequenced the gene for dna ligase from thermus thermophilus. a comparison of this sequence and those of other ligases reveals significant homology only with that of escherichia coli. the overall amino acid composition of the thermophilic ligase and the pattern of amino acid substitutions between the two proteins are consistent with compositional biases in other thermophilic enzymes. we have engineered the expression of the t. thermophilus gene in escherichia coli, and we show ... | 1991 | 1840584 |