| influence of ph in the antigenic composition of bacillus amyloliquefaciens. | | 1976 | 26010 |
| new chromogenic and fluorogenic substrates for pyrrolidonyl peptidase. | l-pyroglutamyl derivatives of p-nitroaniline and 7-amino-4-methylcoumarin were synthesized as new sensitive substrates for pyrrolidonyl peptidase (pyrrolidonecarboxylyl peptidase) from bacillus amyloliquefaciens. their hydrolyses could be followed by conventional colorimetric and fluorometric procedures; i.e., in terms of the increase in absorbance at 410 nm caused by the liberation of p-nitroaniline and the emission at 440 nm after excitation at 370 nm depending on the liberation of 7-amino-4-m ... | 1978 | 26673 |
| purification and characterization of l-pyrrolidonecarboxylate peptidase from bacillus amyloiliquefaciens. | microorganisms capable of producing l-pyrrolidonecarboxylate peptidase l-pyrrolidonyl peptidase, ec 3.4.11.8 were screened and a strain of bacillus amyloliquefaciens was chosen as one of the most potent producers of the enzyme. the enzyme was purified from lysozyme-lysate of the bacterial cells by salting out with ammonium sulfate, adsorption on deae-cellulose, covalent chromatography on pcmb-sepharose and by gel filtration on sephadex g-150. by these procedures, the enzyme was purified about 80 ... | 1978 | 29893 |
| the substrate specificity of pyrrolidone carboxylyl peptidase from bacillus amyloliquefaciens. | | 1979 | 39608 |
| isolation and characterization of a polynucleotide phosphorylase from bacillus amyloliquefaciens. | bacillus amyloliquefaciens bam-2 produces large amounts of extracellular enzymes, and the synthesis of these proteins appears to be dependent upon abnormal ribonucleic acid metabolism. a polynucleotide phosphorylase (nucleoside diphosphate:polynucleotide nucleotidyl transferase) was identified, purified, and characterized from this strain. the purification scheme involved cell disruption, phase partitioning, differential (nh4)2so4 solubilities, agarose gel filtration, and diethylaminoethyl-sepha ... | 1977 | 45489 |
| release of extracellular enzymes from bacillus amyloliquefaciens. | washed-cell suspensions of bacillus amyloliquefaciens secrete significant amounts of the extracellular enzymes alpha-amylase and protease for about 15 min in the almost complete absence of protein synthesis. this apparently represents release of preformed enzyme en route to secretion. the release was independent of energy but was affected by temperature. pulse-labeling experiments showed that newly synthesized enzyme molecules are either immediately released into the external medium or equilibra ... | 1975 | 47325 |
| the restriction endonucleases in bacillus amyloliquefaciens n strain. substrate specificities. | two species of restriction endonuclease were isolated by gel filtration and deae-cellulose chromatography from a cell-free extract of bacillus amyloliquefaciens (b. subtilits) n strain; a lower molecular weight endonuclease (endonuclease r.bamni) and a higher molecular-weight one (endonuclease r.bamnx). both of them required only mg2+ for their activities. endonuclease r.bamnx introduced a larger number of site-specific scissions in excherchia coli phage lambda dna that endonuclease r.bamni did. ... | 1976 | 182257 |
| human papillomavirus dna: physical mapping of the cleavage sites of bacillus amyloliquefaciens (bami) and haemophilus parainfluenzae (hpaii) endonucleases and evidence for partial heterogeneity. | the dna of human papillomavirus (hpv) obtained from a pool of plantar warts is cleaved by bacillus amyloliquefaciens (bami) and haemophilus parainfluenzae (hpaii) restriction endonucleases at one and four specific sites, respectively. these sites were localized on the previously established cleavage map of hpv dna, using the hind, hindiii, hpai, and ecori endonuclease restriction sites as reference. the four hpaii sites were mapped, clockwise, at 1.4, 41.1, 44.3, and 52.8% of the genome length f ... | 1977 | 191644 |
| location of the cleavage sites on the sv 40 dna map produced by the restriction endonucleases pst 1 and bam 1. | restriction endonucleases from providencia stuartii (pst 1) and bacillus amyloliquefaciens h (bam 1) cleave sv 40 dna at two and one specific sites, respectively. using ecori and hind iii endonuclease restriction sites as reference, the two pst i sites were mapped at 0.050; 0.265 and the bam i site was mapped at 0.170 of the genome length, clockwise, from the single ecori cleavage site. | 1978 | 210371 |
| phage lambda receptor chromosomes for dna fragments made with restriction endonuclease i of bacillus amyloliquefaciens h. | | 1979 | 231662 |
| alpha-amylase from five strains of bacillus amyloliquefaciens: evidence for identical primary structures. | the alpha-amylases from five strains of bacillus amyloliquefaciens were compared to determine whether differences in primary structure are responsible for variations in catalytic properties previously reported among the enzymes. amino acid analysis established virtually identical compositions for the proteins. reaction with dimethylaminoaphthylene sulfonylchloride indicated the amino-terminal amino acid of each amylase to be valine. carboxyl termini of the enzymes have been determined by digesti ... | 1978 | 306994 |
| alpha-glucosidase, a membrane-bound enzyme of alpha-glucan metabolism in bacillus amyloliquefaciens. purification and partial characterization. | the organism bacillus amyloliquefaciens is capable of producing alpha-amylase (1,4-alpha-d-glucan glucanohydrolase, ec 3.2.1.1) and isoamylase (glycogen 6-glucanohydrolase, ec 3.2.1.68) extracellurlarly and a membrane-bound, intracellular alpha-glucosidase (alpha-d-glucoside glucohydrolase, ec 3.2.1.20). the amounts of alpha-glucosidase in cells of b. amyloliquefaciens grown on amylaceous polysaccharides were significantly higher then in cells grown on non-carbohydrate carbon sources. alpha-gluc ... | 1978 | 339955 |
| [physicochemical properties of an extracellular polysaccharide isolated from culture media of bacillus amyloliquefaciens]. | a new extracellular polysaccharide has been isolated by chromatography on anion exchanger of a fraction obtained from highly viscous culture media of bacillus amyloliquefaciens. this polysaccharide is characterised by high molecular weight (1,000,000 dalton) and intrinsic viscosity (323 ml/g). it contains 24% neutral sugar (galactose and mannose 5:1), 35% glucuronic acid and 51.5% n-acetylhexosamines (n-actylglucosamine, n-acetylgalactosamine and n-acetylbacillosamine 6:9:1). | 1977 | 413665 |
| physiological responses of bacteria to cytochalasin a: effects on growth, transport, and enzyme induction. | cytochalasin a at 5 to 25 microgram/ml (1.0 x 10(-5) to 5.2 x 10(-5) m) inhibited the growth of three gram-positive bacteria, arthrobacter sialophilus, staphyloccus aureus, and bacillus amyloliquifaciens, but had little or no effect on the growth of three gram-negative bacteria, excherichia coli, pseudomonas maltophilia, and aeromonas proteolytica. a. sialophilus and s. aureus recovered spontaneously from cytochalasin a-mediated growth inhibition after a considerable lag period, which was depend ... | 1979 | 422516 |
| purification, properties and specificity of the restriction endonuclease from bacillus stearothermophilus. | the restriction endonuclease bsti was purified from 70kg of bacillus stearothermophilus. the final product is at least 97% pure as judged by sodium dodecyl sulphate/polyacrylamide-gel electrophoresis; this major protein species co-migrates with the enzyme activity on native polyacrylamide-gel electrophoresis and isoelectric focusing. pure restriction endonuclease bsti has a subunit mol.wt. of 26,000 and is probably a loosely associated dimer. the enzyme shows maximum activity at ph values betwee ... | 1979 | 426781 |
| [isolation and some properties of restriction endonuclease from bacillus amyloliquefaciens]. | a partially purified preparation of restriction endonuclease bam i was isolated from the cells of bacillus amyloliquefaciens. the purification procedure was a modification of a method described by wilson and young. isolation and purification of the enzyme involved disruption of the cells by ultrasonication, treatment with streptomycin sulfate, fractionation by ammonium sulfate, chromatography on deae-cellulose and hydroxylapatite and rechromatography on deae-cellulose. restrictase bam i splits t ... | 1978 | 656508 |
| membrane phospholipid asymmetry in bacillus amyloliquefaciens. | the phospholipid distribution in the membrane of bacillus amyloliquefaciens was studied by using phospholipase c (b. cereus), phospholipase a2 (crotalus), and the nonpenetrating chemical probe trinitrobenzenesulfonic acid. after treatment of intact protoplasts of b. amyloliquefaciens with either phospholipase, about 70% of total membrane phospholipid was hydrolyzed; specifically, about 90, 90, and 30% of phosphatidylethanolamine, phosphatidylglycerol, and cardiolipin, respectively. under these ... | 1978 | 681277 |
| effect of growth temperature on membrane fatty acid composition and susceptibility to cold shock of bacillus amyloliquefaciens. | we investigated the fatty acid composition of the membrane of bacillus amyloliquefaciens grown at different temperatures. a decrease in growth temperature was accompanied by an increase in the ratio of branched- to straight-chain fatty acids and a marked increase in the level of unsaturation of branched-chain fatty acids. when cells of this organism grown at 30 degrees c were cold shocked, viability and ability to secrete extracellular protease were lost. growth of this organism at lower tempera ... | 1978 | 690073 |
| modulation of an apparent mrna pool for extracellular protease in bacillus amyloliquefaciens. | late-log-phase cells of bacillus amyloliquefaciens have the unusual capacity to produce extracellular protease for over 60 min in the presence of rifampin or actinomycin d at levels which strongly inhibit incorporation of amino acids into cellular protein. if cells are incubated in the presence of high levels of amino acids for 75 min this capacity is exhausted, but it is retained if the incubation is carried out in low levels of amino acids. transfer of exhausted cells from high to low concentr ... | 1978 | 711667 |
| sequence specificity of dna methylases from bacillus amyloliquefaciens and bacillus brevis. | | 1978 | 712853 |
| purification and characterization of the sequence-specific endonuclease bam hi. | the specific endonuclease bam hi from bacillus amyloliquefaciens (rub 500) has been purified to apparent homogeneity. two active forms of the enzyme corresponding to the dimeric and tetrameric forms have been isolated. on sodium dodecyl sulfate-polyacrylamide gel electrophoresis, the enzyme dissociated into mr = 22,000 +/- 500 subunits. bam hi has a broad ph optimum on the alkaline side and requires mg2+ which can be partially replaced by mn2+. the enzyme catalysis appears to be governed by a tw ... | 1979 | 762109 |
| messenger ribonucleic acid content of bacillus amyloliquefaciens throughout its growth cycle compared with bacillus subtilis 168. | | 1975 | 809591 |
| evidence for somatic rearrangement of immunoglobulin genes coding for variable and constant regions. | a high-molecular-weight dna from balb/c mouse early embryo or from mopc 321 plasmacytoma (a k-chain producer) was digested to completion with bacillus amyloliquefaciens strain h restriction enzyme (bamh i). the resulting dna fragments were fractionated according to size in preparative agarose gel electrophoresis. dna fragments carrying gene sequences coding for the variable or constant region of k chains were detected by hybridization with purified, 125i-labeled, whole mopc 321 k mrna and with i ... | 1976 | 824647 |
| recognition sequence of specific endonuclease bamh.i from bacillus amyloliquefaciens h. | | 1977 | 834250 |
| complementation of peptides of barnase, extracellular ribonuclease of bacillus amyloliquefaciens. | recovery of ribonuclease activity by complementation of peptides of barnase is reported. activity is restored to barnase-(1-102), which lacks eight amino acids from its cooh terminus, by combination with peptides-(88-110), -(95-110), or -(99-108), and also with succinylated peptide-(88-110). the dissociation constants are about 8 x 10(-6) m for the first two combinations and little, if any, greater for the other two. based on barnase-(1-102) concentration, up to 80% of native activity is obtaine ... | 1977 | 863882 |
| preparation and characterization of a dextran-amylase conjugate. | bacillus amyloliquefaciens alpha-amylase was attached to dextran after activation of the polysaccharide by using a modification of the cyanogen bromide method. the soluble dextran-amylase conjugate was purified by molecular-sieve chromatography. the conjugated enzyme has greater stability than the unmodified enzyme at low ph values, during heat treatment, and on removal of calcium ions with a chelating agent. attachment of dextran to alpha-amylase did not alter the michaelis constant of the enzy ... | 1976 | 963699 |
| subsite mapping of enzymes. application of the depolymerase computer model to two alpha-amylases. | in the preceding paper (allen and thoma, 1976) we developed a depolymerase computer model, which uses a minimization routine to establish a subsite map for a depolymerase. in the present paper we show how the model is applied to experimental data for two alpha-amylases. michaelis parameters and bond-cleavage frequencies for substrates of chain lengths up to twelve glucosyl units have been reported for bacillus amyloliquefaciens, and a subsite map has been proposed for this enzyme [thoma et al. ( ... | 1976 | 999630 |
| location in bacteriophage lamdba dna of cleavage sites of the site-specific endonuclease from bacillus amyloliquefaciens h. | the sites in escherichia coli bacteriophage lambda dna cleaved by the site-specific endonuclease isolated from bacillus amyloliquefaciens h (bami) are found to be at 0.114, 0.466, 0.580, 0.713, and 0.861 lambda units. the sites were located by analysis of the products of digestion of lambda dna and lambda-ara transducing phage dna, and verified by double digestion with bami and ecori. | 1976 | 1083914 |
| stimulation of the differential rate of exoenzyme formation in bacillus amyloliquefaciens by streptolydigin, an inhibitor of rna chain elongation. | | 1975 | 1113077 |
| a two-state conformational transition of the extracellular ribonuclease of bacillus amyloliquefaciens (barnase) induced by sodium dodecyl sulfate. | barnase, the extracellular ribonuclease of bacillus amyloliquefaciens, is shown to undergo a reversible two-state conformational transition at 0.65 mm sodium dodecyl sulfate (sds) aat 37 degrees. the prinicipal evidence is based on the equivalence of two independent values of the sds-barnase binding ratio; about 14 mol of sds/mol of barnase. both were derived from fluorometric titration data, one being based on simple conservation of sds and the other on the use of wyman's theory of linked funct ... | 1975 | 1138866 |
| evidence for the presence of alpha amylase in the cell membrane of bacillus amyloliquefaciens. | | 1975 | 1147980 |
| relationship between exoprotease secretion and the synthesis of ribonucleic acid and protein in bacillus amyloliquefaciens. | studies with washed bacteria suspended in fresh medium, in which bacterial densities were altered by a factor of four so as to cause accelerated entry of exponential bacteria into the postexponential phase and to re-establish growth in postexponential bacteria, have been performed. under all the conditions examined rifampin, at a concentration of 0.5 mug/ml, inhibited [(14)c]uracil incorporation into total ribonucleic acid (rna) by 90 to 95%. the percentage of inhibition of incorporation of (14) ... | 1975 | 1155927 |
| defective bacteriophage lambda chromosome, potential vector for dna fragments obtained after cleavage by bacillus amyloliquefaciens endonuclease (bami). | | 1975 | 1157934 |
| evidence for extrusion of unfolded extracellular enzyme polypeptide chains through membranes of bacillus amyloliquefaciens. | the production of extracellular alpha-amylase and protease by protoplasts of bacillus amyloliquefaciens has been achieved. the production of enzymically active protease was totally dependent on a high concentration of either mg2+, ca2+, or spermidine, but production of active alpha-amylase was not. this cation dependence of protease production was seen immediately upon addition of lysozyme to intact cells. the cations could prevent the inactivation of protease and alter the cytoplasmic membrane ... | 1975 | 1158850 |
| identification of isoamylase, a glycogen-debranching enzyme, from bacillus amyloliquefaciens. | | 1975 | 1175769 |
| stability of rapidly labelled messenger ribonucleic acid in bacillus amyloliquefaciens during the phases of minimum and maximum extracellular enzyme formation. | | 1975 | 1177310 |
| isolation of a sequence-specific endonuclease (bami) from bacillus amyloliquefaciens h. | | 1975 | 1177312 |
| structures of singly branched heptaoses produced by bacterial liquefying alpha-amylase. | 1. a singly branched heptaose produced as a limit dextrin in the digest of beta-limit dextrin with liquefying alpha-amylase [ec 3.2.1.1] of bacillus amyloliquefaciens was isolated in a paper chromatographically pure state. 2. analysis using several enzymes revealed that the isolated branched dextrin was a mixture of six singly branched heptaoses with different ramifying points. 3. all the branched heptaoses contained a 62-alpha-maltosylmaltotriose moiety in their molecules, differing only in the ... | 1975 | 1240101 |
| the effect of rifampicin on the stability of the messenger ribonucleic acid of bacillus amyloliquefaciens as determined by dna:rna hybridization. | rifampicin at a concentration of 10 mug/ml completely inhibited protein synthesis in exponential-phase cultures of bacillus amyloliquefaciens. at this same concentration the drug was shown to have no effect on the stability of mrna, determined as the functional and hybridizable material, when compared with hybridizable mrna in an uninhibited system. in each case, the half-life of the mrna had a value in the range 5 +/- 1 min, at 30 degrees c. | 1976 | 1245839 |
| [cloning and gene expression of bacillus cereus neutral proteinase in bacillus subtilis cells]. | the neutral proteinase gene of bacillus cereus was cloned. its restriction map and the direction of transcription was determined. it was shown that the neutral proteinase gene could be expressed in bacillus cells. the thermostability of the product coded by the neutral proteinase gene and its natural analogue was explored. the obtained data indicate that the neutral proteinase of bacillus cereus is closely related to the enzyme of bacillus amyloliquefaciens by these parameters. it was found that ... | 1992 | 1339956 |
| identification of glutamic acid 105 at the active site of bacillus amyloliquefaciens 1,3-1,4-beta-d-glucan 4-glucanohydrolase using epoxide-based inhibitors. | bacillus amyloliquefaciens 1,3-1,4-beta-d-glucan 4-glucanohydrolase (ec 3.2.1.73) was modified by the mechanism-based, affinity-labeling reagent [14c](3,4)-epoxybutyl beta-d-cellobioside. following partial inactivation a completely inactivated enzyme preparation containing 1.1 mol of covalently bound inhibitor/mol of protein was obtained by chromatography on a cellulosic matrix. the inactivated enzyme was digested with endoproteinase glu-c and radioactive peptides purified by reversed-phase high ... | 1992 | 1360982 |
| cloning and expression of alpha-amylase from bacillus amyloliquefaciens in a stable plasmid vector in lactobacillus plantarum. | lactobacillus plantarum is used in a wide range of agricultural and food fermentations. in this paper we report the introduction of alpha-amylase into the organism from bacillus amyloliquefaciens on a stable recombinant plasmid. the genetically manipulated organism grew on mrsb medium supplemented with starch and it may be a prototype for the development of lactobacilli able to use an increased range of substrates in commercial fermentations. | 1990 | 1366859 |
| genetic manipulation of bacillus amyloliquefaciens. | application of modern gene technology to strain improvement of the industrially important bacterium bacillus amyloliquefaciens is reported. several different plasmid constructions carrying the alpha-amylase gene (amye) from b. amyloliquefaciens were amplified in this species either extrachromosomally or intrachromosomally. the amye gene cloned on a pub110-derived high copy plasmid pkth10 directed the highest yields both in rich laboratory medium and in crude industrial medium. the alpha-amylase ... | 1991 | 1367238 |
| molecular cloning in lactobacillus helveticus by plasmid psa3::pva797 co-integrate formation and conjugal transfer. | a gene encoding beta-glucanase activity from bacillus amyloliquefaciens was subcloned in both orientations into plasmid shuttle vector psa3. in only one orientation could a co-integrate be generated with the conjugative plasmid pva797. the plasmid co-integrate was conjugated into lactobacillus helveticus strain cnrz450, where it was stably maintained without antibiotic selection and exhibited beta-glucanase activity. this method of introducing cloned dna into thermophilic lactobacilli will facil ... | 1991 | 1367540 |
| processing of the prepropeptide portions of the bacillus amyloliquefaciens neutral protease fused to bacillus subtilis alpha-amylase and human growth hormone during secretion in bacillus subtilis. | a set of nested 3'-terminal deletions of the prepropeptide of the bacillus amyloliquefaciens neutral protease gene was constructed. alpha-amylase and human growth hormone were secreted using these truncated genes in bacillus subtilis. the level of the secreted alpha-amylase varied with the region for the truncated prepropeptide contained in the fusion gene but was independent of its length. even though length of the prepropeptide varied, the mobilities of secreted alpha-amylases were the same as ... | 1992 | 1367948 |
| effect of immobilisation on the production of alpha-amylase by an industrial strain of bacillus amyloliquefaciens. | the effect of immobilisation of an industrial strain of bacillus amyloliquefaciens in calcium alginate beads on production of alpha-amylase was investigated using lactose-based media in shake flasks and in a 0.3 dm3 glass fermenter. although the microorganism was a good alpha-amylase producer in batch cultures of free cells, it was unable to produce the enzyme for extended periods either in repeated batch cultures, or in continuous cultivation. in each case, parallel tests with cells immobilised ... | 1992 | 1368003 |
| secretion of correctly processed and folded pancreatic secretory trypsin inhibitor by bacillus subtilis. | we constructed a plasmid, designated pnpp126, containing a dna sequence encoding a fusion protein composed of bacillus amyloliquefaciens neutral protease prepeptide (signal peptide) and human pancreatic secretory trypsin inhibitor (hpsti), where the mature hpsti is accurately fused to the 3'-terminal of the prepeptide coding region. it was observed that the strain bacillus subtilis mt600 harboring pnpp126 could secrete a trypsin inhibitory activity into the culture medium. the n-terminal amino a ... | 1992 | 1368060 |
| expression of bacillus amyloliquefaciens amylase and vibrio alginolyticus protease a fusion genes. | previously we reported [deane, s. m., maharaj, r., robb, f. t. & woods, d. r. (1987) journal of general microbiology 133, 2295-2302] that the production of a vibrio alginolyticus sds-resistant alkaline serine protease (pro a) cloned in escherichia coli was characterized by a 12 h delay between the synthesis of an inactive precursor and secretion of active pro a. replacement of the v. alginolyticus promoter region by the alpha-amylase promoter region from bacillus amyloliquefaciens resulted in th ... | 1992 | 1373436 |
| positive theta-angles in proteins by nuclear magnetic resonance spectroscopy. | non-glycine residues with positive theta-angles have been identified in four proteins, barley serine proteinase inhibitor ci-2, bacterial ribonuclease (barnase) of bacillus amyloliquefaciens, hen egg white lysozyme and a basic protein from barley seed (barwin) by use of nuclear magnetic resonance spectroscopy. by accurate measurements of the coupling constant (3)jhnhalpha and integration of the nuclear overhauser hn-halpha cross peak, positive theta-angles could be determined reliably to 60 degr ... | 1992 | 1392567 |
| dna sequence and analysis of a cryptic 4.2-kb plasmid from the filamentous cyanobacterium, plectonema sp. strain pcc 6402. | the 4194-bp plasmid, prf1, from plectonema sp. strain pcc 6402 was completely sequenced and analyzed. seven potential open reading frames were identified. the predicted amino acid sequence of open reading frame c (orf c) had identities of 34, 29, and 25% with rep b from the staphylococcus aureus plasmid, pub110; rep from the bacillus amyloliquefaciens plasmid, pftb14; and protein a from the s. aureus plasmid, pc194, respectively. a 75-amino-acid region conserved in these proteins (rep b, rep, an ... | 1992 | 1409974 |
| synthesis and secretion of an erwinia chrysanthemi pectate lyase in saccharomyces cerevisiae regulated by different combinations of bacterial and yeast promoter and signal sequences. | nine different expression-secretion cassettes, comprising novel combinations of yeast and bacterial gene promoters and secretion signal sequences, were constructed and evaluated. a pectate lyase-encoding gene (pele) from erwinia chrysanthemi was inserted between each one of these expression-secretion cassettes and a yeast gene terminator, generating recombinant yeast-integrating shuttle plasmids pams1 through pams9. these yip5-derived plasmids were transformed and stably integrated into the geno ... | 1992 | 1427097 |
| regulation of the bamhi restriction-modification system by a small intergenic open reading frame, bamhic, in both escherichia coli and bacillus subtilis. | bamhi, from bacillus amyloliquefaciens h, is a type ii restriction-modification system recognizing and cleaving the sequence g--gatcc. the bamhi restriction-modification system contains divergently transcribed endonuclease and methylase genes along with a small open reading frame oriented in the direction of the endonuclease gene. the small open reading frame has been designated bamhic (for bamhi controlling element). it acts as both a positive activator of endonuclease expression and a negative ... | 1992 | 1429443 |
| microbial degradation of shrimp-shell waste. | a total of 40 strains of bacteria were isolated and tested for their potentiality to degrade chitin and utilize shrimp-shell waste for the production of chitinase. the activity ratio, percentage of weight loss and enzyme activity were determined for cultures exhibiting highest chitinolytic activities. the most active organisms were identified as alcaligenes denitrificans, bacillus amyloliquefaciens, b. megaterium and b. subtilis. the potentiality of the first two organisms to degrade chitin was ... | 1992 | 1512701 |
| a positive selection vector for cloning high molecular weight dna by the bacteriophage p1 system: improved cloning efficacy. | the bacteriophage p1 cloning system can package and propagate dna inserts that are up to 95 kilobases. clones are maintained in escherichia coli by a low-copy replicon in the p1 cloning vector and can be amplified by inducing a second replicon in the vector with isopropyl beta-d-thiogalactopyranoside. to overcome the necessity of screening clones for dna inserts, we have developed a p1 vector with a positive selection system that is based on the properties of the sacb gene from bacillus amyloliq ... | 1992 | 1549564 |
| translocation mediated by domain ii of pseudomonas exotoxin a: transport of barnase into the cytosol. | pseudomonas exotoxin a (pe) is a protein toxin composed of three structural domains. functional analysis of pe has revealed that domain i is the cell-binding domain and that domain iii functions in adp ribosylation. domain ii was originally designated as the translocation domain, mediating the transfer of domain iii to the cytosol, because mutations in this domain result in toxin molecules with normal cell-binding and adp-ribosylation activities but which are not cytotoxic. however, the results ... | 1992 | 1567815 |
| modular expression and secretion vectors for bacillus subtilis. | a modular vector system has been developed for the extracellular production of heterologous proteins in bacillus subtilis. this modular vector system consists of four secretion vectors which are based upon the genes encoding the bacillus amyloliquefaciens extracellular alkaline protease, neutral protease, barnase and levansucrase. the modular vectors contain compatible restriction sites downstream from the signal peptide-coding region. three reporter proteins (staphylococcal protein a, levansucr ... | 1992 | 1587474 |
| extensive comparison of the substrate preferences of two subtilisins as determined with peptide substrates which are based on the principle of intramolecular quenching. | subtilisins are serine endopeptidases with an extended binding cleft comprising at least eight binding subsites. interestingly, subsites distant from the scissile bond play a dominant role in determining the specificity of the enzymes. the development of internally quenched fluorogenic substrates, which allow polypeptides of more than 11 amino acids to be inserted between the donor and the acceptor, has rendered it possible to perform a highly systematic mapping of the individual subsites of the ... | 1992 | 1627543 |
| expression of the erwinia carotovora polygalacturonase-encoding gene in bacillus subtilis: role of signal peptide fusions on production of a heterologous protein. | the peha gene encoding an endopolygalacturonase (pectinase) of erwinia carotovora subsp. carotovora has been cloned previously [saarilahti et al., mol. microbiol. 4 (1990) 1037-1044]. we expressed peha in bacillus subtilis using a secretion vector based on the promoter and signal sequence of the alpha-amylase (amy)-encoding gene, amye, from bacillus amyloliquefaciens. to test whether the location of the junction between the secretion vector and peha affects the protein yield, we made four differ ... | 1992 | 1628841 |
| [isolation and analysis of protease-deficient mutants of bacillus amyloliquefaciens]. | four types of protease negative mutants of bacillus amyloliquefaciens a50 were selected after four stages of step-by-step uv light mutagenesis. edta, and pmsf were used as inhibitors of protease activity to characterize the protease negative mutants with regard to the protease type. the electrophoretic patterns of proteases from culture medium of b. amyloliquefaciens protease deficient mutants were studied. the proinsulin stability in the culture medium of different mutants was analysed. proteas ... | 1992 | 1639263 |
| pathway and stability of protein folding. | we describe an experimental approach to the problem of protein folding and stability which measures interaction energies and maps structures of intermediates and transition states during the folding pathway. the strategy is based on two steps. first, protein engineering is used to remove interactions that stabilize defined positions in barnase, the rnase from bacillus amyloliquefaciens. the consequent changes in stability are measured from the changes in free energy of unfolding of the protein. ... | 1991 | 1678536 |
| levansucrase: a tool to study protein secretion in bacillus subtilis. | the bacillus amyloliquefaciens levansucrase gene (sacb[bamp]) was engineered in such a way that a heterologous gene could be inserted between the second and third codon of the mature levansucrase. extracellular levansucrase activity was detected only when the heterologous protein was secreted into the growth medium. a positive selection system to isolate suppressors of signal sequence mutants in bacillus subtilis has been developed based on the secretion of levansucrase. | 1991 | 1784817 |
| sequence regions of bacilli metalloproteinases that can affect enzyme thermostability. | by a computer analysis of the five bacilli metalloprotease sequences it was found that mesophilic bacillus amyloliquefaciens and b. subtilis proteases had lost two ca(2+)-binding sites due to the substitutions asp----ser 57, asp----thr 59, asp----pro 200, in comparison with the thermostable b. thermoproteolyticus thermolysin and b. stearothermophilus protease, which conserved three ca(2+)-binding sites, and b. cereus protease with the intermediate thermostability, which had presumably lost only ... | 1991 | 1812491 |
| optimization of bacillus alpha-amylase production by saccharomyces cerevisiae. | production of bacillus amyloliquefaciens alpha-amylase by saccharomyces cerevisiae using the multicopy plasmid paah5 and ways of improving the yields of secreted enzyme were studied. in standard non-buffered medium, alpha-amylase was rapidly inactivated but stabilization of the ph at 6 led to stable accumulation of alpha-amylase in the culture medium. removal of 1100 bp of the upstream sequence of the adh1 promoter present on paah5 resulted in delayed but increased alpha-amylase production: 29-f ... | 1991 | 1872026 |
| effect of signal sequence alterations on export of levansucrase in bacillus subtilis. | a series of alterations in the bacillus amyloliquefaciens levansucrase signal peptide were made by in vitro mutagenesis, and their effect on the secretion of levansucrase in bacillus subtilis was studied. some of the alterations resulted in a completely defective signal peptide. these included the removal of positively charged residues from the n-terminus and disruption of the hydrophobic core of the signal peptide either by introducing a charged residue or by deleting five or more amino acids. ... | 1991 | 1898923 |
| barnase toxin: a new chimeric toxin composed of pseudomonas exotoxin a and barnase. | we have constructed a chimeric toxin composed of pseudomonas exotoxin a (pe) and the extracellular ribonuclease of bacillus amyloliquefaciens, barnase. the chimeric protein, termed pe-bar, reacted with both anti-pe and anti-barnase antisera and had both adp ribosylation and ribonuclease activities. the chimeric toxin was cytotoxic to the murine fibroblast cell line l929 and to a murine hybridoma resistant to pe. a mutant form of pe-bar lacking adp-ribosylating activity was still cytotoxic to l92 ... | 1991 | 1900455 |
| use of alkaline phosphatase fusions to study protein secretion in bacillus subtilis. | we have constructed a vector designed to facilitate the study of protein secretion in bacillus subtilis. this vector is based on a translational fusion between the expression elements and signal sequence of bacillus amyloliquefaciens alkaline protease and the mature coding sequence for escherichia coli alkaline phosphatase (phoa). we show that export of alkaline phosphatase from b. subtilis depends on a functional signal sequence and that alkaline phosphatase activity depends upon secretion. the ... | 1991 | 1901054 |
| optimization of the signal-sequence cleavage site for secretion from bacillus subtilis of a 34-amino acid fragment of human parathyroid hormone. | we have effected the secretion from bacillus subtilis of a 34-amino acid (aa) fragment of human parathyroid hormone (pth,1-34), using a bacillus amyloliquefaciens neutral protease signal sequence. the secretion efficiency depended on the aa sequence near the signal-sequence cleavage site. we constructed a series of gene fusions encoding different pairs of aa between the signal sequence and pth,1-34. there was a correlation between those polypeptides which were efficiently secreted and the potent ... | 1991 | 1908402 |
| accurate measurements of coupling constants from two-dimensional nuclear magnetic resonance spectra of proteins and determination of phi-angles. | a new and simple method to measure 3jhnh alpha coupling constants of proteins by adding and subtracting traces from corresponding two-dimensional nuclear overhauser enhanced spectroscopy and two-dimensional correlated spectroscopy cross peaks after scaling is proposed. the optimal scaling for the addition and the subtraction of the two traces is obtained by minimizing an error function. the method was proven to give accurate and precise measurements of coupling constants when tested with a serie ... | 1991 | 2005622 |
| hybrid bacillus (1-3,1-4)-beta-glucanases: engineering thermostable enzymes by construction of hybrid genes. | hybrid (1-3,1-4)-beta-glucanase genes were constructed by extension of overlapping segments of the (1-3,1-4)-beta-glucanase genes from bacillus amyloliquefaciens and b. macerans generated by the polymerase chain reaction (pcr). four hybrid genes were expressed in escherichia coli cells. the mature hybrid enzymes contain a 16, 36, 78, or 152 amino acid n-terminal sequence derived from b. amyloliquefaciens (1-3,1-4)-beta-glucanase followed by a c-terminal segment derived from b. macerans (1-3,1-4) ... | 1991 | 2005860 |
| aromatic-aromatic interactions and protein stability. investigation by double-mutant cycles. | the side-chains of phenylalanine and tyrosine residues in proteins are frequently found to be involved in pairwise interactions. these occur both within repeating elements of secondary structure and in tertiary and quaternary interactions. it has been suggested that they are important in protein folding and stability, and non-bonded potential energy calculations indicate that a typical aromatic-aromatic interaction has an energy of between -1 and -2 kcal/mol and contributes between -0.6 and -1.3 ... | 1991 | 2010920 |
| crystal structure of a barnase-d(gpc) complex at 1.9 a resolution. | the ribonuclease excreted by bacillus amyloliquefaciens, barnase, was co-crystallized with the deoxy-dinucleotide d(gpc). the crystal structure was determined by molecular replacement from a model of free barnase previously derived by mauguen et al. refinement was carried out using data to 1.9 a resolution. the final model, which has a crystallographic r factor of 22%, includes 869 protein atoms, 38 atoms from d(gpc), a sulfate ion and 73 water molecules. only minor differences from free barnase ... | 1991 | 2023257 |
| overexpression, purification and crystallization of bamhi endonuclease. | the type ii restriction endonuclease bamhi has been expressed in e. coli, producing 100-fold more enzyme than the wild type bacillus amyloliquefaciens h strain. this high yield has facilitated purification to homogeneity of large amounts of the enzyme, along with its crystallization in a form which diffracts to at least 1.9 a in x-ray analysis. | 1991 | 2030964 |
| m.h2i, a multispecific 5c-dna methyltransferase encoded by bacillus amyloliquefaciens phage h2. | bacillus amyloliquefaciens phage h2 codes for a multispecific cytosine-5-dna- methyltransferase (mtase), m.h2i, which methylates ggcc, gcngc and [sequence: see text] target sequences. the gene coding for m.h2i was cloned in escherichia coli and its nucleotide (nt) sequence was determined. it consists of 1509 bp, corresponding to a protein of 503 amino acids (aa) with a calculated mr of 57,166. a comparison of the aa sequence of m.h2i with those of the multispecific mtases encoded by bacillus sub ... | 1991 | 2055471 |
| co-expression of a saccharomyces diastaticus glucoamylase-encoding gene and a bacillus amyloliquefaciens alpha-amylase-encoding gene in saccharomyces cerevisiae. | a glucoamylase-encoding gene (sta2) from saccharomyces diastaticus and an alpha-amylase-encoding gene (amy) from bacillus amyloliquefaciens were cloned separately into a yeast-integrating shuttle vector (yip5), generating recombinant plasmids psp1 and psp2, respectively. the sta2 and amy genes were jointly cloned into yip5, generating plasmid psp3. subsequently, the dominant selectable marker aph1, encoding resistance to geneticin g418 (gtr), was cloned into psp3, resulting in psp4. for enhanced ... | 1991 | 2055483 |
| production and secretion of pertussis toxin subunits in bacillus subtilis. | pertussis toxin (pt) is a major component of today's acellular whooping cough vaccines. the use of acellular vaccines is predicted to increase sharply in the near future. there is therefore a need to produce pt in a way that makes its purification as easy as possible. our approach was to express all five pt subunits individually in bacillus subtilis. we have used vectors containing the promoter and signal sequences of the alpha-amylase gene of bacillus amyloliquefaciens followed by an insert enc ... | 1990 | 2110091 |
| bacillus amyloliquefaciens alpha-amylase signal sequence fused in frame with human proinsulin is properly processed by bacillus subtilis cells. | the plasmid pbins1, containing the promoter, sd and leader peptide sequences of bacillus amyloliquefaciens alpha-amylase gene and 267 bp long sequence coding for human proinsulin directs the efficient synthesis of hybrid preproinsulin, as well as quantitative secretion of proinsulin outside of protease-deficient bacillus subtilis aj73 cells. the recombinant proinsulin has been isolated from the culture medium and its n-terminal sequence shown to be identical with that of natural human prohormone ... | 1990 | 2112381 |
| excretion into the culture medium of a bacillus beta-glucanase after overproduction in escherichia coli. | the beta-glucanase gene (bgl) from bacillus amyloliquefaciens was expressed in e. coli csh 55 under the control of the pr promoter of phage lambda that is repressed by the thermosensitive repressor c1857. production of beta-glucanase was drastically stimulated by a temperature shift to 42 degrees c. this overexpression of the bgl gene (about 20% of the total cellular protein) led to an almost complete excretion of the otherwise periplasmic protein into the extracellular medium, beta-glucanase ac ... | 1990 | 2114118 |
| efficiency of transcriptional terminators in bacillus subtilis. | to promote more efficient synthesis of heterologous gene products in a bacillus subtilis host, we have developed a system for rapidly testing the effect of a putative terminator on in vivo gene expression. terminator structures from the bacillus amyloliquefaciens amye gene, the bacillus licheniformis penp gene, the b. subtilis bgls gene, and the bacillus thuringiensis cry gene were subcloned and inserted into a vector in such a way as to disrupt expression of the cat-86 gene. comparisons are mad ... | 1990 | 2123811 |
| secretion of the alpha-galactosidase from cyamopsis tetragonoloba (guar) by bacillus subtilis. | a fusion of dna sequences encoding the spo2 promoter, the alpha-amylase signal sequence from bacillus amyloliquefaciens, and the mature part of the alpha-galactosidase from cyamopsis tetragonoloba (guar) was constructed on a bacillus subtilis multicopy vector. bacillus cells of the protease-deficient strain db104 harboring this vector produced and secreted the plant enzyme alpha-galactosidase up to levels of 1,700 u/liter. a growth medium suppressing the residual proteolytic activity of strain d ... | 1990 | 2160224 |
| the complete sequence of the bacillus amyloliquefaciens strain h, cellular bamhi methylase gene. | | 1990 | 2235511 |
| isolation and characterization of levansucrase-encoding gene from bacillus amyloliquefaciens. | the gene encoding levansucrase (lvs) from bacillus amyloliquefaciens (sacb[bamp]) was isolated, sequenced and expressed in bacillus subtilis. analysis of the nucleotide sequence of sacb[bamp] reveals extensive homology with that of the b. subtilis lvs-encoding gene in the promoter and coding region. the sacb[bamp] gene cloned in a multicopy plasmid is induced by sucrose in b. subtilis. | 1990 | 2265762 |
| an investigation of the cause of the eosinophilia-myalgia syndrome associated with tryptophan use. | the eosinophilia-myalgia syndrome is a newly recognized illness that has been associated with the consumption of tryptophan products. it is not known whether the cause is related to the tryptophan itself or to chemical constituents introduced by the manufacturing process. | 1990 | 2370887 |
| the complete sequence of the bacillus amyloliquefaciens proviral h2, bamhi methylase gene. | | 1990 | 2374727 |
| dna hybridization procedure to detect pseudorabies virus dna in swine tissue. | a dna hybridization technique was developed to detect the presence of pseudorabies virus (prv) dna. p nick translated probes of high specific activity were prepared from transformed escherichia coli plasmids into which bacillus amyloliquefaciens h (bam h1) restriction fragments of prv dna had been inserted. swine cellular dna and tissue culture prv dna were digested with bam h1, separated by agarosegel electrophoresis, transferred onto nitrocellulose paper, hybridized to the radioactive probes, ... | 1985 | 2408522 |
| [limited proteolysis of human leukocyte interferon-alpha2 and localization of the antigenic determinant binding the monoclonal antibodies]. | large peptide fragments of human leucocyte interferon-alpha 2 (inf-alpha 2) were obtained by limited proteolysis with trypsin, pepsin, thermolysine and bacillus amyloliquefaciens intracellular serine proteinase. the ability of the fragments to bind murine monoclonal antibodies nk2 raised against inf-alpha 2 was studied by the immunoblotting technique. the region of sequence 110-149 is the most sensitive to proteolytic attack, being probably exposed on the surface of the inf-alpha 2 molecule. inf ... | 1985 | 2415171 |
| sigma 29-like protein is a common sporulation-specific element in bacteria of the genus bacillus. | a monoclonal antibody specific for an antigenic determinant on the bacillus subtilis sporulation-induced sigma factor sigma 29 reacted with proteins similar in size to sigma 29 in extracts of sporulating bacillus licheniformis, bacillus amyloliquifaciens, bacillus cereus, bacillus natto, and bacillus pumilus but not in extracts prepared from vegetatively growing cultures of these bacteria. these results indicate that rna polymerase modifications, initially described for b. subtilis, are likely t ... | 1985 | 2415506 |
| complete nucleotide sequence of a gene coding for heat- and ph-stable alpha-amylase of bacillus licheniformis: comparison of the amino acid sequences of three bacterial liquefying alpha-amylases deduced from the dna sequences. | the gene coding for the heat-stable and ph-stable alpha-amylase of bacillus licheniformis 584 (atcc 27811) was cloned in escherichia coli and the nucleotide sequence of a dna fragment of 1,948 base pairs containing the entire amylase gene was determined. as inferred from the dna sequence, the b. licheniformis alpha-amylase had a signal peptide of 29 amino acid residues and the mature enzyme comprised 483 amino acid residues, giving a molecular weight of 55,200. the amino acid sequence of b. lich ... | 1985 | 2418011 |
| tunicamycin-resistant mutants of bacillus amyloliquefaciens are deficient in amylase, protease and penicillinase synthesis and have altered sensitivity to antibiotics and autolysis. | mutants of bacillus amyloliquefaciens resistant to at least 10 micrograms/ml of tunicamycin were isolated and shown to be pleiotropic. the mutants were more resistant to streptomycin, chloramphenicol, kanamycin and neomycin than was the parent strain but less resistant to penicillin g and tetracycline. they were more autolytic, presumably due to an altered cell wall. the mutants produced reduced levels of amylase, penicillinase and both metal and serine protease besides having an enhanced sporul ... | 1986 | 2424885 |
| [two-codon mutagenesis of alpha-amylase gene of bacillus amyloliquefaciens]. | the oligonucleotide encoding bam hi recognition site having the structure pcgggatc had been inserted into the recognition sites mspi of the b. amyloliquefaciens alpha-amylase gene, which was cloned in ptg29b plasmid. the alpha-amylase gene had no bamhi sites before mutagenesis. the set of pnsbamhi plasmids with bamhi site at four different positions was obtained. it was shown that all the mutant alpha-amylases possess different specific activities. one of the mutant proteins possesses reduced th ... | 1988 | 2464738 |
| on the mechanism of growth of cells (bacillus amyloliquefaciens) in the mixed aqueous two-phase system. | the growth of bacillus amyloliquefaciens in the aqueous two-phase system, made up of polyethylene glycol, dextran, and water, was investigated. generally, bacillus partitions in the dextran phase, but the magnitude of the separation depends largely on the overall composition of polymers in the phase system. the kinetics of growth of bacillus amyloliquefaciens was studied in the polyethylene glycol-rich continuous phase, dextran-rich dispersed phase, and in the mixed phase. from the kinetic data ... | 1989 | 2472777 |
| expression of bovine pancreatic ribonuclease a coded by a synthetic gene in bacillus subtilis. | two different hybrid genes were constructed which fuse the bacillus amyloliquefaciens alkaline protease gene (apr[bamp]) promoter and signal peptide coding region to a synthetic bpr gene coding for the mature bovine pancreatic rnase a. the first gene fusion (apr-bpr1) contained the apr[bamp] signal peptide coding region fused to mature bpr through a linker coded 3-amino acid region and retained the signal processing site ala-ala of the alkaline protease. the second fusion (apr-bpr2) joined the e ... | 1989 | 2501158 |
| production of pneumolysin, a pneumococcal toxin, in bacillus subtilis. | we have expressed the pneumolysin gene of streptococcus pneumoniae in bacillus subtilis, both from its own promoter and as a fusion protein. the level of expression of pneumolysin from its own promoter was low. the protein produced was hemolytically active. a higher level of expression (about 10 micrograms/ml of culture) was achieved when either one of two c-terminal fragments (corresponding to amino acids 265-471 and 55-471, respectively) or the entire coding part of the pneumolysin gene were f ... | 1989 | 2502471 |
| protein engineering of disulfide bonds in subtilisin bpn'. | five single-disulfide mutants were studied in subtilisin bpn', a cysteine-free, secreted serine protease from bacillus amyloliquefaciens. the disulfides were engineered between residues 26-232, 29-119, 36-210, 41-80, and 148-243. these bonds connected a variety of secondary structural elements, located in buried or exposed positions at least 10 a from the catalytic ser-221, and linked residues that were separated by 39 up to 206 amino acids. all disulfide bonds formed in the enzyme when the expr ... | 1989 | 2504281 |
| [cloning the alpha-amylase gene of bacillus amyloliquefaciens in cyanobacteria cells]. | the recombinant plasmids of piah4amy series were constructed containing the alpha-amylase gene of bacillus amyloliquefaciens a50 with its own promoter and leading sequence within an integrative vector plasmid piah4 (cmr) for cyanobacterium anacystis nidulans r2. at anacystis nidulans transformation the hybrid plasmids integrate into cyanobacterium chromosome with high efficiency and all cmr transformants produce alpha-amylase. expression of bacillar alpha-amylase gene in cyanobacterium cells is ... | 1989 | 2515431 |
| transformation of bacillus amyloliquefaciens by electroporation. | a method for transformation of whole bacillus amyloliquefaciens cells by electroporation was developed. the procedure is as efficient as the protoplast transformation method, resulting in up to 10(5) transformants/micrograms plasmid dna, but requires less effort and time. cells for electroporation were grown to late exponential phase in a rich medium supplemented with 0.25 m sucrose, washed with and resuspended in 0.25 m sucrose, 1 mm hepes, 1 mm mgcl2, 10% (v/v) glycerol, ph 7.0, at 3-5 x 10(10 ... | 1989 | 2599355 |
| kinetic characterization of the recombinant ribonuclease from bacillus amyloliquefaciens (barnase) and investigation of key residues in catalysis by site-directed mutagenesis. | barnase, the ribonuclease from bacillus amyloliquefaciens, has been cloned and expressed in escherichia coli [hartley, r. w. (1988) j. mol. biol. 202, 913-915], thus enabling the overproduction and site-directed mutagenesis of one of the smallest enzymes (mr equals 12,382). as barnase is also composed of just a single polypeptide chain with no disulfide bridges and has a reversible folding transition, it affords a fine system for studying protein folding and design. we show here that the recombi ... | 1989 | 2665810 |
| synthesis and secretion of bacterial alpha-amylase by the yeast saccharomyces cerevisiae. | alpha-amylase from bacillus amyloliquefaciens, synthesized in yeast saccharomyces cerevisiae without substitution of the signal sequence, is efficiently secreted from yeast cells: 60-70% of the overall amount of the enzyme is found in the culture fluid. in contrast to many yeast secretory proteins, which accumulate in the periplasmic space and in the cell wall, intracellular alpha-amylase is localized mainly in the cytoplasm. obviously, transfer across the cell wall is not a rate-limiting step i ... | 1989 | 2666166 |
| hybrid bacillus endo-(1-3,1-4)-beta-glucanases: construction of recombinant genes and molecular properties of the gene products. | hybrid beta-glucanase genes were constructed by the reciprocal exchange of the two halves of the isolated beta-glucanase genes from bacillus amyloliquefaciens and b. macerans. the beta-glucanase hybrid enzyme 1 (h1) contains the 107 amino-terminal residues of mature b. amyloliquefaciens beta-glucanase and the 107 carboxyl-terminal amino acid residues of b. macerans beta-glucanase. the reciprocal beta-glucanase hybrid enzyme 2 (h2) consists of the 105 amino-terminal residues from the b. macerans ... | 1989 | 2673278 |
| barnase and barstar: two small proteins to fold and fit together. | barnase and barstar are the extracellular ribonuclease and its intracellular inhibitor produced by bacillus amyloliquefaciens. both are small single-chain proteins and thus are suitable for application to the study of how a protein's sequence directs its fold. barnase has neither disulfide bonds nor non-peptide components and unfolds reversibly in what closely approximates a two-state reaction. the genes for both these proteins have been cloned in e. coli. expression of barstar is necessary to c ... | 1989 | 2696173 |
| investigation into the nature of a bacillus promoter cloned into a promoter-probe plasmid. | the alpha-amylase-coding gene (amy) of bacillus amyloliquefaciens ncp1 was cloned into the bacillus subtilis promoter probe vector ppl603b.1, using a bglii digest of chromosomal dna. the resulting plasmid, pvc102, was shown to have a bglii site within the insert. it was determined that this was the result of the fortuitous co-cloning of 2.88-kb and 0.92-kb bglii fragments separated in ncp1 dna by approx. 3 kb. unexpectedly, this co-cloning was readily repeated. subcloning showed that while the 2 ... | 1989 | 2806909 |