翻译起始区突变对猪卵透明带-3β蛋白在E.coli表达的影响  

Effect of TIR Modification on Expression of pZP3β in E.coli

作　　者：吴健虹[1] 陈小佳[1] 朱伟杰[2] 谢秋玲[1] 孙奋勇[1] 徐万祥 李菁[4] 洪岸[1] 

机构地区：[1]暨南大学生物工程研究所,广州510632 [2]暨南大学生殖免疫研究中心,广州510632 [3]上海市计划生育科研所,上海200032 [4]暨南大学医学院病理生理教研室,广州510632

出　　处：《生殖与避孕》2005年第10期579-587,616,共10页Reproduction and Contraception

基　　金：广东省科技计划项目(2001C12001)

摘　　要：目的:探讨翻译起始区(translation initiation region,TIR)突变对两种氨基酸序列长度不同的猪卵透明带蛋白pZP3β161和pZP3β263在大肠杆菌中表达的影响。方法:应用DNASIS软件辅助分析,分别设计5条不同的上游引物对两种pZP3β基因的TIR区的G+C含量和自由能进行合理调整,PCR扩增后将获得的基因片段克隆入pET-3c载体,于大肠杆菌BL21(DE3)pLysS中诱导表达。结果:TIR对pZP3β表达有明显的影响,其表达的pZP3β161蛋白和pZP3β263蛋白分别占菌总蛋白的25％和15％。结论:在不改变氨基酸的条件下,通过对目的基因TIR进行适当改造,能够有效提高目的蛋白的表达效率。Objective： To obtain two different porcine zona pellucida proteins （pZP3β） in amino acid length, pZP3β161 and pZP3β263, and to further study whether the translation initiation region （TIR） effects expression efficiency of exogenous gene in E.coli. Methods： Five modified upstream primers and one downstream primer were designed for each truncated protein by DNASISv2.5 software and then were employed to adjust G＋C content and A GO of the TIR. Using pET-3c as vector, the cloned pZP3[3161 and pZP3β263 were expressed in BL21（DE3） pLysS after induced. Results： Expression efficiency ofpZP3β protein was strongly influenced by TIR. pZP3β161 and pZP3β263 highly expressed up to 25% and 15%, respectively. Conclusion： Modification of TIR without changing amino acid sequence is an effective tool to enhance the expression efficiency of desired protein.

关 键 词：猪卵透明带 截短型 翻译起始区(TIR) 基因表达 

分 类 号：Q78[生物学—分子生物学]