【摘要】 根据已发表的Bacilluslicheniformis植酸酶基因DNA序列(GeneBankAF469936),设计并合成了一对引物,应用PCR技术,以地衣芽孢杆菌AB91062的总DNA为模板,扩增出了植酸酶基因phyL的编码区,并将其克隆到pMD18-T载体上,进行了序列测定,测序结果经Genetool序列分析表明该基因编码区长1146bp,编码381个氨基酸,与已发表的Bacilluslicheniformis植酸酶基因序列在核苷酸水平上与所发表的序列有92%的同源性;在氨基酸水平上与已发表的衣芽孢杆菌植酸酶相似性为93%.并将该基因编码区克隆到大肠杆菌表达质粒pHBM625上,并在大肠杆菌中实现了高效表达,表达产物具有正常的生物学活性.更多还原

【Abstract】 According to the published phytase gene sequence from Bacillus licheniformis,primers were designed to amplify phytase gene by PCR.The PCR product was cloned to pMD18-T and sequenced.The phytase gene we cloned comprises 1?146 nucleotides and encodes a polypeptide of 381 amino acids.Compared with published phyL gene sequence,the phytase gene showed 92?% identity on nucleotides level and 93?% identity on amino acids level. It was also cloned to the expression vector pHBM625 and overexpressed in E.coli.The expressed product has normal bioactivity.更多还原