芽孢杆菌植酸酶基因的克隆及生物信息学分析

采用PCR技术从Bacillus amyloliquefaciens DSM 1061中克隆到植酸酶的全长基因phyC(1152bp)，并将该基因成功克隆到表达载体pET22b(+)，经PCR鉴定和DNA测序证明，pET22b(+)-phyC重组质粒构建成功。将该酶的氨基酸序列在Swiss-prot数据中进行BLAST比对发现，该序列未见报道，其与来自Bacillus subtilis的植酸酶序列(Swiss-prot ID: O31097)最相似，序列一致性为98.7%。分析可知该氨基酸序列在位点为128、144、153、257和370处分别变异为Ala、Ile、Ser、Pro和Lys。运用SignalP 3.0服务器对其信号肽进行预测，表明该酶N端1～26个残基可能是信号肽序列。运用SWISS-MODEL服务器进行同源建模，经PROCHECK V3.5软件对其Ramachandran plot、G-factor等进行评价，结果显示结构模型中的键长、键角及二面角的分布及结构合理。该基因已在GenBank上登录(登录号为HM747163)。

Gene Cloning and Bioinformatics Analysis of Phytase from Bacillus amyloliquefaciens

The full-length DNA sequence of neutral phytase gene was cloned from Bacillus amyloliquefaciens DSM 1061 by PCR,and successfully inserted into the expression vector pET22b(+).The results were identified by PCR and DNA sequencing.Based on Blast alignment,it was found that the amino acid sequence of the enzyme has never been reported the Swiss-prot database.Its amino acids sequence showed 98.7% identity with that of Bacillus subtilis(Swiss-prot ID:O31097).The residues 128,144,153,257 and 370 are replaced by A...