非融合膜外型猪卵透明带-3β蛋白在原核系统中的表达  

Expression of Nonfusion Extracellular Porcine Zona Pellucida Protein 3β in E.coli

作　　者：陈小佳[1] 谢秋玲[1] 朱伟杰[2] 孙奋勇[1] 徐万祥 洪岸[1] 李菁[4] 吴健虹[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5期259-262,299,共5页Reproduction and Contraception

基　　金：广东省科技计划项目(2001C12001)

摘　　要：目的:通过基因工程的方法在原核表达系统中表达去除了信号肽和跨膜区的膜外型猪卵透明带蛋白pZP3β。方法:设计适宜引物,采用PCR方法,在基因水平对膜外型pZP3β的翻译起始序列进行适当调整,获得的基因克隆入pET-3c载体,并进一步在大肠杆菌E.coli BL21(DE3) pLysS中表达。结果:得到表达率约10%的非融合膜外型pZP3β蛋白的表达,并经Western-blot鉴定为目的蛋白。结论:外源蛋白在不改变氨基酸序列条件下,在基因水平的适当调整可以有效提高其表达率。Objective: To obtain the recombinant nonfusion extracellular procine zona pellucida protein 3β (pZP3β). Methods: pZP3 cDNA with a modified nucleotide sequence of initiation translation was obtained by PCR using a pair of approptiate primers and then was cloned into pET-3c vector. The positive reconstitute plasmid was transformed into the host, E.coli BL21(DE3) pLysS. Results: The recombinant nonfusion extracellular pZP3β was expressed in E.coli to 10% of total cellular proteins, and identified by Western-blot. Conclusion: Modification of nucleotide sequence without changing amino acid sequence is an effective means to increase nonfusion expression level of recombinant proteins, such as pZP3β in E.coli.

关 键 词：非融合 膜外型 猪卵透明带 基因表达 

分 类 号：Q819[生物学—生物工程]