梅花鹿促卵泡激素α亚基基因克隆与融合表达

Cloning and Fusion Expression of FSHα-subunit Gene in Cervus Nippon

以原构建的克隆载体为模板,PCR扩增梅花鹿FSHα亚基基因,TA克隆后经双酶切插入表达载体pGEX-6P-2,阳性克隆导入E.coli BL21,IPTG诱导表达GST-FSHα融合蛋白,SDS-PAGE进行分析鉴定。结果表明,FSHαPCR产物大小约380bp,测序结果与GenBank序列一致;重组表达载体pGEX-6P-2-FSHα构建成功;融合蛋白经SDS-PAGE分析,结果发现,在分子质量39.3ku处出现特异性蛋白质条带,说明梅花鹿FSHα亚基基因片段已在E.coli BL21中成功表达了FSHα-GST融合蛋白。

FSHα-subunit gene segments were amplified by PCR from the original constructed vector.Then the PCR product was TA cloned,sequenced and inserted into the carrier pGEX-6P-2.The recombinant plasmids were identified by restricted enzymes digestion,and then the positive clones were transformed into E.coli BL21.GST-FSHα fusion proteins were expressed via the induction of IPTG,and detected by SDS-PAGE.The results showed that PCR product was about 380 bp,and sequencing result was identical with GenBank.The pGEX-6P-2-FSHα positive clone produced a 39.3 ku fusion protein on SDS-PAGE gel.Successfully cloning and expressing Cervus nippon FSHα-subunit gene.