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[目的]对铜绿假单胞菌脂肪酶Lipase基因进行原核表达。[方法]利用PCR方法从铜绿假单胞菌基因组DNA中扩增得到脂肪酶基因,测定其核苷酸序列,利用基因重组技术构建脂肪酶基因的原核表达载体,加IPTG至终浓度为1.0 mmol/L诱导蛋白表达4 h,并进行SDS-PAGE电泳。[结果]从铜绿假单胞菌中克隆的脂肪酶基因成熟肽的序列,与NCBI上所递交的铜绿假单胞菌脂肪酶序列同源性很高,达99.36%。成功构建了脂肪酶基因的原核表达载体pET32a-Lip,进一步SDS-PAGE电泳结果显示目的基因得到高效表达。[结论]克隆的铜绿假单胞菌脂肪酶带有自身的信号肽,也可以在大肠杆菌中正常表达,可以用于进一步的研究。 相似文献
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铜绿假单胞菌脂肪酶Lipase基因的原核表达(英文) 总被引:4,自引:1,他引:3
[Objective] The aim of this study was to investigate the prokaryotic expression of pseudomonas aeruginosa Lipase gene.[Method]Lipase gene was amplified by PCR from the genome DNA of pseudomonas aeruginosa,and its nucleotide sequence was determined.The prokaryotic expression vector of Lipase gene was constructed by the gene recombination technique.The protein expression was induced for 4 hours by IPTG with the final concentration of 1.0 mmol/L,and then SDS-PAGE electrophoresis was analyzed.[Result]The sequence of mature peptides in Lipase gene cloned from pseudomonas aeruginosa had a 99.36% homology with that of pseudomonas aeruginosa lipase submitted in NCBI,so the prokaryotic expression vector of Lipase gene pET32a-Lip was successfully constructed.Furthermore,the results of SDS-PAGE electrophoresis showed that the target gene was expressed highly and effectively.[Conclusion]The cloned pseudomonas aeruginosa lipase with its signal peptide could be normally expressed in E.coli and also used for further study. 相似文献
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