人干扰素INF-ω在植物乳杆菌中的表达与鉴定

Expression and identification of human IFN-ω in Lacbacillus plantarum

目的以植物乳杆菌为载体,将干扰素IFN-ω基因导入植物乳杆菌内,作为先期探索和尝试,构建重组阴道微生态菌株。方法采用抗菌肽诱导双组分调节表达系统,以细菌素Sakacin A为诱导物、sapK和sapR为双组分调节基因的穿梭质粒pSIP300,将干扰素ω基因置于pSIP300上,构成重组质粒pSIP300-ω,在大肠埃希菌DH5α中进行扩增,分别电转化3株植物乳杆菌:Lacbacillus plantarum1.3,Lacbacillus plantarum1.11,Lacbacillusplantarum14917。结果与结论经SDS-PAGE与ELISA检测证实,成功构建了能够表达IFN-ω重组植物乳杆菌Lacbacillus plantarum1.11/pSIP300-ω。

Objective To construct recombinant vaginal strain by using Lactobacillus plantarum as the interferon-ω delivering vector. Method SapR and sapK belong to the family of bacterial two-component signal transduction systems. Biotin sakacin A was used as the inducer,SapK and sapR were used as the cognate regulatory system. IFN-ω was linked in the two-component signal transduction shuttle vector pSIP300. The recombinant vector pSIP300-ω was electrotransferred to Lacbacillus plantarum 1.3 , Lacbacillus plantarum 1.11 ,and Lacbacillus plantarum 14917 respectively. Result and conclusion and conclusion By SDS-PAGE and ELISA detecting,a recombinant/xwbac//lus p/antarum 1. I 1/pSIP300-ωexpressing human IFN-ω was successfully constructed.