植物乳杆菌ZS2058的亚油酸异构酶基因在乳酸克鲁维酵母中的克隆表达

目的：将植物乳杆菌ZS2058（Lactobacillus plantarum ZS2058）的亚油酸异构酶基因在乳酸克鲁维酵母（Kluyveromyces lactis）中进行克隆表达。方法：根据NCBI中已报道亚油酸异构酶（linoleate isomerase，LAI）基因的序列特征，设计引物对筛得的植物乳杆菌ZS2058进行PCR扩增，得到亚油酸异构酶全基因序列，克隆至乳酸克鲁维酵母表达载体pKLAC1，电转化得重组菌pKLAC1-LAI /Kluyveromyces lactis GG799。结果：SDS-PAGE检测，重组菌进行分泌表达获得目的蛋白，大小约为67 kDa；气相色谱（Gas Chromatogram，GC）检测到共轭亚油酸（conjugated linoleic acids，CLA）典型峰。结论：植物乳杆菌ZS2058中的亚油酸异构酶基因在乳酸克鲁维酵母中得到分泌表达，重组酶转化效率约为26%。

Cloning and Expression of the Linoleate Isomerase Gene from Lactobacillus plantarum ZS2058 in Kluyveromyces lactis GG799

Objective:To express the Linoleate isomerase gene of Lactobacillus plantarum in Kluyveromyces lactis.Method:Linoleate isomerase (LAI)gene was amplified by PCR from chromosome of Lactobacillus plantarum ZS2058,then the gene was cloned into Kluyveromyces lactis expression vector pKLAC1,the recombinant plasmid pKLAC1-LAI was transformed into Kluyveromyces lactis GG799 by electroporation,The expression level and the activity of recombinant enzyme were detected by SDS-PAGE and Gas Chromatogram.Result:It was demo...