荣昌猪白细胞介素-10(IL-10)基因的克隆及原核表达  

Cloning and Prokaryolic Expression of Rongchang Pig Porcine IL-10

作　　者：李晓琪[1] 郭万柱[1] 黄亚平[1] 张丽[1] 

机构地区：[1]四川农业大学动物生物技术中心,四川雅安625014

出　　处：《安徽农业科学》2007年第15期4446-4447,4459,共3页Journal of Anhui Agricultural Sciences

基　　金：国家"973"项目(2004CCA01800)

摘　　要：根据GenBank收录的猪白细胞介素-10(PIL-10)设计1对特异性引物,经刀豆蛋白素A(conA)诱导猪淋巴细胞并提取总RNA,RT-PCR方法首次扩增出荣昌猪PIL-10的全长基因。序列分析表明PIL-10全长771 bp。设计1对引物亚克隆荣昌猪IL-10基因成熟蛋白编码基因(477 bp)。利用基因重组技术构建了PET32a(+)-PIL-10融合表达载体,经酶切鉴定,DNA测序证实重组质粒构建正确。将重组质粒转化大肠杆菌BL21,IPTG诱导,SDS-PAGE,重组菌的RT-PCR,Western blotting结果表明融合表达成功。The eDNA sequence encoding Rongchang pig porcine intertonkin- 10（PIL- 1 ） was cloned from the lymphocyte stimulated with Con A by means of the specific primers based on the reported sequence in GonBank.Sequence identification indicated the full length of cloned cDNA was 771 bp.Another pair of primers was designed to sub-clone gone coding porcine IL-10 mature protein.The PE132a（ ＋ ）-PIL-10 was identified and analyzed with enzymatic digestion and DNA sequcing and was confirmed.The recombinant strain was transformed into E. coli BL21 ,induced by IPTG,SDS-PAGE and the RT-PCR result of recombinant strain confirmed PIL-10 was presented successfully.

关 键 词：荣昌猪 白细胞介素-10 克隆 原核表达 

分 类 号：Q78[生物学—分子生物学]