来航鸡FOXL2基因的克隆与原核表达  

Cloning and prokaryotic expression of gene coding FOXL2 of leghorn

作　　者：徐萍[1] 李任峰[1] 赵坤[1] 鲁毅[1] 王三虎[1] 

机构地区：[1]河南科技学院动物科学学院,河南新乡453003

出　　处：《中国兽医学报》2013年第4期622-626,共5页Chinese Journal of Veterinary Science

基　　金：河南省基础与前沿重点研究资助项目(082300430020)

摘　　要：根据GenBank已发表的红色原鸡FOXL2基因序列(登录号NM_001012612.1)设计引物,以来航鸡全血为模板,PCR扩增出其基因全长编码区,经BamHⅠ-HindⅢ双酶切后定向克隆于pET28a原核表达载体,获得pET28a-FOXL2重组原核表达载体。将携带有重组原核表达载体的大肠杆菌BL21(DE3)通过0.5mmol/L IPTG进行诱导表达,经过SDS-PAGE电泳检测,显示诱导表达蛋白大小大约为37 000,与预期表达蛋白大小一致。Western blot-ting检测显示该蛋白为His融合蛋白,表明重组原核表达载体在大肠杆菌中成功表达出了目的融合蛋白。为进一步探索该基因的生物学功能奠定了基础。A pair of primers was designed based on the published nudeotide sequence of the Gallus FOXL2 gene （GenBank accession number：NM_001012612.1）. The full-length coding sequence of the leghorn FOXL2 DNA was amplified by PCR using a template of the leghorn whole blood. The product was double-digested by Barn H I and Hind III, and then directionally cloned into the pET28a vector. The recombinant expression plasmid in the E. coli BL21 （DE3） was induced with 0.5 mmol/L IPTG. SDS-PAGE analysis showed that the expressed protein was about 37 000. Western blotting revealed that the expressed protein was the His tag fusion protein, which indicated that the recombinant prokaryotic expression vector expressed the fusion protein successfully. The cloning and expression of the leghorn FOXL2 gene lays the foundation for further study on its biological function.

关 键 词：来航鸡 FOXL2基因 克隆 原核表达 

分 类 号：S831.2[农业科学—畜牧学]