安徽农业科学2011，Vol.39Issue(10)：5898-5900,3.
犬IL-2基因克隆及原核表达
Cloning and Prokaryotic Expression of Canine IL-2 Gene
摘要
Abstract
[ Objective ] To clone and express canine IL-2 gene and thus to provide theoretical support for the development of novel immune enhancers and genetic engineering vaccines. [ Method] Leukocytes separated from canine whole blood were stimulated by concanavalin for 20 h ,and then total RNA was extracted. According to the sequence of canine IL-2 gene published in the GenBank,a pair of primers was designed. After PCR amplification,the target fragment was cloned into prokaryotic expression vector pET-28a. The recombinants were transformed into the host bacteria BL21. After IPTG induction,the expression products were analyzed by SDS-PAGE. [ Result] A 500 bp band with the expected size appeared in the RT-PCR products. After the pMD18-T-IL2 was identified by double dignstion,an approximately 500 bp fragment was produced,which indicated successful cloning of the gene. After the pET-28a-IL2 was identified by restriction enzyme digestion and PCR,a 500 bp fragment was produced,which. indicated successful construction of the expression vector. As revealed by the SDS-PAGE analysis,a protein band with molecular weight of about 20 kDa appeared. [ Conclusion] The canine IL-2 gene was cloned and expressed.关键词
犬/白细胞介素 2/基因克隆/原核表达Key words
Canine/ Interleukin-2/ Gene cloning/ Prokaryotic expression分类
农业科技引用本文复制引用
费东亮,毕聪明,苏禹刚,白刃..犬IL-2基因克隆及原核表达[J].安徽农业科学,2011,39(10):5898-5900,3.基金项目
辽宁省教育厅资助项目(L2010263). （L2010263）
