嗜酸乳杆菌的亚油酸异构酶基因克隆及其pMG36e表达载体的构建

以嗜酸乳杆菌基因组为模板,对亚油酸异构酶基因进行PCR,PCR产物克隆到pMD-19T质粒中,经菌落PCR、酶切分析和DNA测序鉴定克隆成功后,亚克隆入乳酸菌表达质粒pMG36e,构建乳酸菌表达载体pMG36e-LAI并转化到大肠杆菌DH5α中,经菌落PCR和酶切分析初步证明表达载体pMG36e-LAI构建成功。该研究为利用工程菌工业化生产高产量、高活性亚油酸异构酶制剂来生产CLA奠定基础。

The cloning of LAI gene and the construction of pMG36e expression vector

Employing the genomic DNA of Lactobacillus acidophilus strain as the template of PCR,then subcloning the PCR product into the pMD-19T vector,we sucessfully finished the endonuclease analysis and the cloning procedure after sequencing the coding region. Furthermore,we cut the above PCR product and pMG36e with the same restriction enzyme,with the purpose of getting the engineering strain of Lactobacillus MG1363 pMG36e-LAI. The construction of this new recombinant expression vector will lay the base of the study on obtaining large and high active linolenic acid isomerase and developing the new microbial ecological agent.