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木糖还原酶(XR, EC 1.1.1.21)是真菌微生物代谢木糖的关键酶之一。本文以米曲霉基因组DNA为模板,克隆木糖还原酶基因(xr,GenBank登录号:FJ957890.1),并对XR的序列、系统进化树、理化性质及蛋白结构等进行生物信息学分析。结果表明: xr基因序列长1449 bp,其中开放阅读框长960 bp,编码319个氨基酸,蛋白质分子质量35.9 kDa,等电点为5.78;米曲霉XR与其他菌种XR有较高的同一性,含有醛酮还原酶家族的两个指纹结构和一个参与辅酶结合活性位点指纹结构,以及醛酮还原酶家族典型的(β/α)8 TIM桶结构,说明米曲霉XR属于醛酮还原酶家族。 相似文献
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We previously reported the DNA sequence and expression of
the Chlorobium vibrioforme glutamyl-tRNA reductase (hemA)
gene (Majumdar et al., Arch Microbiol 156:281, 1991). The sequence downstream
of the hemA gene indicated homology to Escherichia coli and
Bacillus subtilis porphobilinogen deaminase (hemC) gene. The
Chlorobium gene was confirmed to be the porphobilinogen deaminase
gene, and complete sequence of the structural gene was obtained. A 2.8-kb DNA
fragment containing the 1.3-kb hemA gene of Chlorobium was
cloned into a hemC auxotroph (Sz16) of Bacillus subtilis, and
complementation of the auxotroph to prototrophy was achieved. DNA sequence
data showed a single open reading frame of 840 bp coding a protein of 279
amino acid residues. The deduced amino acid sequence of the
Chlorobium porphobilinogen deaminase revealed 39% to
46% homology with the corresponding prokaryotic and eukaryotic
sequences.
Received: 20 September 1996 / Accepted: 26 October 1996 相似文献
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大蒜花叶病毒外壳蛋白基因cDNA的克隆和序列分析 总被引:1,自引:0,他引:1
我们从自然发病的大蒜中分离得到了大蒜花叶病毒。以其基因组RNA为模板合成了3'末端部分cDNA。从中选出一批插入片段在2.0kb以上的重组克隆,经Northern点杂交分析证实了所选克隆与基因组RNA同源。通过对若干个克隆的插入片段两端部分序列的测定,选出一个克隆pGM495,其插入片段的长度约为2.4kb,3′末端存有一个Poly(A)结构,它应包含了编码该病毒外壳蛋白全部序列。序列测定的结果表明,这个cDNA片段全长为2379bp,其中含有与酶切图谱分析结果相符的EeoRI、PstI及BamHI酶切位点。第一个终止密码子TAA与3′g末端相距264bp,我们根据碱基序列推定的氨基酸序列与其它已发表的Potyvirus的外壳蛋白氨基酸序列以及外壳蛋白翻译后加工的蛋白酶专一切点相比较后推测,编码该病毒外壳蛋白序列可能起始于3′末端上游的1170bp处,共编码302个氨基酸,其分子量为36kD,略大于SDS-PAGE所测定的33kD,非编码区域长264bp,富含AT,并有多个终止密码子的存在。趾3′末端32~27bp处有一个AATAA序列。 相似文献
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以嗜水气单胞菌BZ和NK分离株的DNA为模板, 采用PCR技术, 扩增气溶素基因(aerA)的DNA片段, 将其克隆到pMD18-T载体上。通过序列测定, 分析结果表明:所克隆的1393 bp片段为aerA部分序列, 编码产生464个氨基酸。BZ与NK之间aerA核苷酸同源性为97.6%, 氨基酸同源性为98.3%, 与其它分离物核苷酸同源性为71.6%~97.5%, 氨基酸同源性为68.0%~98.9%。利用邻接法构建了aerA分子树状图, 树状图分析表明:气单胞菌属各分离物聚为三支, 其中嗜水气单胞菌各菌株之间关系密切, 被聚类为同一支。 相似文献
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嗜水气单胞菌气溶素基因的克隆与序列分析 总被引:2,自引:0,他引:2
以嗜水气单胞菌BZ和NK分离株的DNA为模板,采用PCR技术,扩增气溶素基因(aerA)的DNA片段,将其克隆到pMDl8-T载体上.通过序列测定,分析结果表明:所克隆的1393 bp片段为aerA部分序列,编码产生464个氨基酸.BZ与NK之间aerA核苷酸同源性为97.6%.氨基酸同源性为98.3%,与其它分离物核苷酸同源性为71.6%~97.5%,氨基酸同源性为68.0%~98.9%.利用邻接法构建了aerA分子树状图,树状图分析表明:气单胞菌属各分离物聚为三支,其中嗜水气单胞菌各菌株之间关系密切,被聚类为同一支. 相似文献
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为探究esdC基因在谢瓦氏曲霉间型变种的结构特征,从已构建的抑制性差减文库(SSH)中筛选得到esdC基因的EST序列,采用cDNA末端快速扩增(RACE)技术扩增该基因片段的5’和3’端,拼接得到全长cDNA1472bp。通过序列分析,该cDNA序列含有一个开放阅读框(ORF),长度为819bp,从396~1214bp,含有30个腺嘌呤尾巴。预测编码273个氨基酸,该蛋白质的等电点pI为9.51,分子量为30.26558kD,为不稳定蛋白。序列同源性分析表明:该基因与费希新萨托菌(Neosartorya fischeri NRRL181)有性发育蛋白EsdC相似度最高,为78%,其保守区序列主要集中于蛋白质的N端。本实验为进一步研究esdC基因的分子功能及可能参与的调控途径奠定基础。 相似文献
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《Bioscience, biotechnology, and biochemistry》2013,77(7):1684-1687
Mycodextranase (EC 3.2.1.61) is an α-glucanase that cleaves α-1,4-bonds of alternating α-1,3- and α-1,4-linked D-glucan (nigeran). The gene encoding mycodextranase from Streptomyces sp. J-13-3 was cloned by hybridization with a degenerate oligonucleotide probe from the amino-terminal amino acid sequence of the enzyme and its nucleotide structure was analyzed. The open reading frame consisted of 1,803 base pairs encoding a signal peptide of 60 amino acids and a mature protein of 540 amino acids with a calculated molecular weight of 56,078. The deduced amino acid sequence showed weak similality to a chitinase homolog from Streptomyces lividans and a chitinase from Xanthomonas sp. 相似文献
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Cloning, Nucleotide Sequence, and Expression of the Gene Encoding the Bacteriocin Boticin B from Clostridium botulinum Strain 213B 下载免费PDF全文
Boticin B is a heat-stable bacteriocin produced by Clostridium botulinum strain 213B that has inhibitory activity against various strains of C. botulinum and related clostridia. The gene encoding the bacteriocin was localized to a 3.0-kb HindIII fragment of an 18.8-kb plasmid, cloned, and sequenced. DNA sequencing revealed the boticin B structural gene, btcB, to be an open reading frame encoding 50 amino acids. A C. botulinum strain 62A transconjugant containing the HindIII fragment inserted into a clostridial shuttle vector expressed boticin B, although at much lower levels than those observed in C. botulinum 213B. To our knowledge, this is the first demonstration and characterization of a bacteriocin from toxigenic group I C. botulinum. 相似文献
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《Bioscience, biotechnology, and biochemistry》2013,77(6):1056-1062
We cloned and sequenced the glutathione reductase gene (gor) of an oxygen-tolerant Streptococcus mutans, and constructed a gor-disruption mutant by homologous recombination. The gor gene consisted of 1,350 bp, coding for a protein of 450 amino acid residues. The deduced amino acid sequence of the S. mutans gor gene product showed extensive similarity with those of glutathione reductases from prokaryotes and eukaryotes. Although the mutant could grow aerobically, it showed no growth in the presence of 2 mM diamide, a thiol-specific oxidant. In contrast, growth of the wild-type strain was not significantly inhibited by 2 mM diamide, and glutathione reductase activity was increased 2.2-fold under these conditions. In addition, the level of glutathione reductase activity in the wild-type strain was increased 3.6-fold upon exposure to air, and the elevated level of the enzyme was retained throughout the aerobic growth. Thus, glutathione reductase may be important in protection of S. mutans against oxidative stress. 相似文献
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Inderjeet Kaur Gurvinder Singh Kocher V. K. Gupta 《Indian journal of microbiology》2012,52(4):630-637
Bacillus circulans MTCC 7906, an extracellular alkaline protease producer was genetically characterized. B. circulans genomic DNA was isolated, oligonucleotide primers specific to alkaline protease gene of B. circulans were designed and its PCR amplification was done. The purified PCR product and pTrcHisA vector were subjected to restriction digestion with NcoI and HindIII and transformed into Escherichia coli DH5-α competent cells. The recombinant expression of alkaline protease gene studied by inducible expression and analysis by SDS-PAGE, established that the alkaline protease protein had an estimated molecular size of 46 kDa. Gene sequencing of the insert from selected recombinant clone showed it to be a 1329 bp gene encoding a protein of 442 amino acids. The sequence was blasted and aligned with known alkaline protease genes for comparison with their nucleotide and amino acid sequences. This identified major matches with three closely related subsp. of B. subtilis (B. subtilis subsp. subtilis strain 168, B. subtilis BSn5 and B. subtilis subsp. spizizenii strain W23). The insert also showed a number of substitutions (mutations) with other sp. of Bacillus which established that alkaline protease of B. circulans MTCC 7906 is a novel gene. The phylogenetic analysis of alkaline protease gene and its predicted amino acid sequences also validated that alkaline protease gene is a novel gene and the same has been accessioned in GenBank with accession number . JN645176.1相似文献
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《Bioscience, biotechnology, and biochemistry》2013,77(10):1852-1857
The ATF2 gene, which encodes alcohol acetyltransferase II (AATase II), was cloned from Saccharomyces cerevisiae Kyokai No. 7 (sake yeast). The ATF2 gene coded for a protein of 535 amino acid residues with a calculated molecular mass of 61,909 daltons. The deduced amino acid sequences of the ATF2 showed 36.9% similarity with that of ATF1, which encodes AATase I. The hydrophobicity profiles for the Atf2 protein and Atf1 protein were similar. A transformant carrying multiple copies of the ATF2 gene had 2.5-fold greater AATase activity than the control, and this activity was not significantly inhibited by linoleic acid. A Southern analysis of the yeast genomes in which the ATF2 gene was used as a probe showed that S. cerevisiae and brewery lager yeast have one ATF2 gene, while S. bayanus has no similar gene. 相似文献
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Molecular Cloning and Nucleotide Sequence of the Superoxide Dismutase Gene and Characterization of Its Product from Bacillus subtilis 总被引:1,自引:0,他引:1 下载免费PDF全文
Takashi Inaoka Yoshinobu Matsumura Tetsuaki Tsuchido 《Journal of bacteriology》1998,180(14):3697-3703
Bacillus subtilis was found to possess one detectable superoxide dismutase (Sod) in both vegetative cells and spores. The Sod activity in vegetative cells was maximal at stationary phase. Manganese was necessary to sustain Sod activity at stationary phase, but paraquat, a superoxide generator, did not induce the expression of Sod. The specific activity of purified Sod was approximately 2,600 U/mg of protein, and the enzyme was a homodimer protein with a molecular mass of approximately 25,000 per monomer. The gene encoding Sod, designated sodA, was cloned by the combination of several PCR methods and the Southern hybridization method. DNA sequence analysis revealed the presence of one open reading frame consisting of 606 bp. Several putative promoter sites were located in the upstream region of sodA. The deduced amino acid sequence showed high homology with other bacterial manganese Sods. Conserved regions in bacterial manganese Sod could also be seen. The phenotype of double mutant Escherichia coli sodA sodB, which could not grow in minimal medium without supplemental amino acids, was complemented by the expression of B. subtilis sodA. 相似文献
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《Bioscience, biotechnology, and biochemistry》2013,77(9):1455-1460
The gene encoding an alkaline serine protease from alkaliphilic Bacillus sp. 221 was cloned in Escherichia coli and expressed in Bacillus suhtilis. An open reading frame of 1,140 bases, identified as the protease gene was preceded by a putative Shine-Dalgarno sequence (AGGAGG) with a spacing of 7 bases. The deduced amino acid sequence had a pre-pro-peptide of 111 residues followed by the mature protease comprising 269 residues. The alkaline protease from alkaliphilic Bacillus sp. 221 had higher homology to the protease from alkaliphilic bacilli (82.1% and 99.6%) than to those from neutrophilic bacilli (60.6—61.70/0). Also Bacillus sp. 221 protease and other protease from alkaliphilic bacilli shared common amino acid changes and 4 amino acid deletions that seemed to be related to characteristics of the enzyme of alkaliphilic bacilli when compared to the proteases from neutrophilic bacilli. 相似文献
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马铃薯卷叶病毒外壳蛋白基因的合成,分子克隆和全序列分析 总被引:4,自引:0,他引:4
施一Shen 《Virologica Sinica》1992,7(4):432-435
以分离自马铃薯主栽品种“紫花白”的马铃薯卷叶病毒(PLRV)分离物的RNA为模板,用人工合成的引物,用反转录和随后PCR扩增的方法合成了PLRV外壳蛋白(CP)基因的cDNA,并克隆于pUC19中。进一步用限制酶切和核苷酸序列分析表明,合成的cDNA由627个核苷酸组成(包括起始和终止密码),序列中有Hinc Ⅱ和BamH Ⅰ两个酶切位点,与国外报道一致。和国外的4个PLRV分离物CP基因序列对比结果,具有高度同源性,其同源率达99.0—99.7%。 相似文献