构建表达IL-10的重组乳酸杆菌  

Construction of a recombinant lactobacillus strain expressing IL-10

作　　者：邱志兵[1] 钟良[1] 陈佳婕[1] 戎兰[1] 陈坚[1] 

机构地区：[1]复旦大学附属华山医院消化科,上海200040

出　　处：《复旦学报（医学版）》2011年第2期163-167,共5页Fudan University Journal of Medical Sciences

基　　金：上海市自然科学基金项目(08ZR1402100)

摘　　要：目的构建表达IL-10的重组乳酸杆菌。方法人工合成信号肽系列后与IL-10基因连接后克隆到干酪乳酸杆菌(Lactobacillus casei,L.casei)整合型表达载体pIlac的lac启动子下游,得到重组质粒pIlac-sp-IL10,电穿孔转化L.caseiCECT 5276。挑选转化后的耐药菌落在含红霉素的培养基中培养,抽提染色体DNA,PCR及测序鉴定证实IL-10基因是否整合到L.casei的基因组中,Western blot检测重组蛋白表达。结果 IL-10基因整合到干酪乳酸杆菌的基因组中;重组L.casei产生的目的蛋白部分分泌进入培养上清。结论 IL-10在L.caseiCECT 5276中得到了分泌表达。Objective To construct a recombinant lactobacillus strain expressing IL-10. Methods A signal peptide sequence（sp） was synthesized and cloned into the integrative plasmid pIlac down-stream of lactose-inducible promoter.The recombined plasmid pIlac-sp-IL10 was then introduced into Lactobacillus casei（L.casei）CECT 5276 by electroporation.PCR with genomic DNA of the recombinant lactobacillus as the template was performed to confirm the integration of IL-10 gene into the chromosome.Western blot was used to detect the expression of the recombinant protein. Results The IL-10 gene had been integrated into the genome of L.casei,and the objective protein was partially secreted into the supernatant. Conclusions IL-10 was expressed by recombinant L.casei CECT 5276.

关 键 词：干酪乳酸杆菌 IL-10 信号肽 重组质粒 目的蛋白 

分 类 号：Q785[生物学—分子生物学] Q343.37