β1,3-1,4-葡聚糖酶基因的克隆与序列分析

摘要

Based on the sequence alignment of Bacillus amyloliquefaciens, two pairs of primers were designed and synthesized in this study. Using the total DNA of Bacillus amyloliquefaciens as template, the gene of β-1,3-1,4-glucanase (bgl) was amplified by PCR, which has about 750 bp in length. The gene was cloned into pTG19-T Easy vector, and the recombinant plasmid was identified by PCR, restriction enzyme analysis and sequencing. The homology analysis revealed that the nucleotide sequences of the cloned β-1, 3-1, 4-glucanase gene similar to the B. Amyloliquefacines(M15674),B. Subtilis(D00518) and B. Licheniformis(AY365256) were 99%, 95% and 94%, respectively.%依据GenBank中收录的解淀粉芽孢杆菌(Bacillus am ylolique aciens)β-1,3-1,4-葡聚糖酶基因序列,设计特异性引物,以提取的解淀粉芽孢杆菌基因组DNA为模板,利用聚合酶链式反应(PCR)扩增获得758 bp的β-1,3-1,4-葡聚糖酶基因(bgl),将此目的基因克隆至pTG19-T Easy 载体中,经PCR、限制性内切酶鉴定和克隆片段的序列测定、比较,结果表明该克隆片段扩增准确、可靠.序列比较发现,此片段与解淀粉芽孢杆菌(B.amylolique acines,M15674)、枯草芽孢杆菌(B.subtilis,D00518)和地衣芽孢杆菌(B.licheni formis,AY365256)分别有99％、95％和94％的同源性.