肿瘤坏死因子α单链抗体基因的优化及衣藻叶绿体表达载体的构建  

Optimizing of the Tumor Necrosis Factor Alpha ScFv Gene and Construction of the Chlamydomonas Chloroplast Expression Vector

作　　者：费小雯[1] 周玉娇[2] 邓晓东[2] 

机构地区：[1]海南医学院理学院,海南海口571101 [2]中国热带农业科学院热带生物技术研究所,海南海口571101

出　　处：《热带农业科学》2012年第11期71-76,共6页Chinese Journal of Tropical Agriculture

基　　金：国家自然科学基金(No.30860028);海南省科技计划重点项目(No.061012)

摘　　要：通过在EMBL数据库中搜索鼠源TNF-α单链抗体基因同源性最高的人胚系抗体基因TNF-α-ScFv,进行人源化改造,并选择衣藻叶绿体偏爱的密码子对TNF-α-ScFv基因进行优化,随后构建带psbA启动子:TNF-α-ScFv::psbA 3′终止子表达框的中间载体,再将psbA启动子:TNF-α-ScFv::psbA 3′终止子表达框插入衣藻叶绿体表达载体p322中,通过PCR和酶切鉴定,确定已成功构建了TNF-α-ScFv基因的衣藻叶绿体表达载体。Tumor necrosis factor alpha （TNF-α） is a cytokine with strong biological activity. An over expression of TNF-α can lead to the progression of the inflammatory condition, septic shock and related diseases. Anti-TNF-α antibody can counteraction various harmful effects produced by high concentration of TNF-α in vivo, which has broad clinical application prospects. In this study, murine TNF-α-ScFv was humanized, and then optimized to suit the C. reinhardtii chloroplast codon bias. An intermediate vector containing psbA promoter： TNF-α-ScFv： psbA 3＇ terminator was constructed and subcloned into C. reinhardtii chloroplast expression vector. This laid a good foundation for expressing TNF-α-ScFv in C. reinhardtii chloroplast.

关 键 词：肿瘤坏死因子Α 单链抗体 莱茵衣藻 叶绿体 基因表达 

分 类 号：R73-3[医药卫生—肿瘤]