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Molecular Cloning, Sequence Analysis and Prokaryotic Expression of Ovine Activin Receptor Type IIB(ActRIIB) Gene被引量:1
2014年
Objective] This study aimed to clone ovine activin receptor type llB (Ac-tRIIB) gene, construct the prokaryotic expression vector and express the target gene in vitro, thus providing basis for further function verification. [Method] The template cDNA which was reversely transcribed from total RNA of sheep liver tissue, was subjected to polymerase chain reaction (PCR) using specific primers of ActRIIB. The ful-length cDNA of ovine ActRIIB was obtained by pMD18-T cloning and sequencing for bioinformatics analysis. Ovine ActRIIB encoding sequence was subcloned into prokaryotic expression vector pET41a with restriction sites BamHl/Notl, and then transformed into BL21 (DE3). The induced products by lPTG were analyzed with SDS-PAGE and Western Blot. [Result] The amplified ful-length cDNA of ovine Ac-tRllB gene was 1 564 bp in length (Genbank accession number: JX422071.1) with an open reading frame of 1 539 bp, encoding 512 amino acides. Ovine ActRllB shared the highest homology (99.6%) with bovine ActRllB. ActRllB had highly ho-mologous C-terminal domains and belonged to the TGFβ family. After prokaryotic expression, an approximately 92 kD His-tagged ActRllB recombinant protein was obtained, which was consistent with the excepted result. [Conclusion] Cloning and successful expression of ovine ActRIIB laid solid foundation for further investigation of its biological function.
张雪梅安静张宁刘明军
关键词:SHEEP
激活素受体ⅡB(ActRⅡB)及突变体对绵羊成肌细胞生长分化的作用
激活素受体ⅡB (ActRⅡB)属于TGF-β超家族成员的丝氨酸/苏氨酸蛋白激酶型受体,起着配体结合和特异性识别的双重功能.其突变体与MSTN结合后形成的复合物不能与Ⅰ型受体结合,从而阻断MSTN下游信号传导,抑制MST...
张雪梅安静刘明军
关键词:肉用绵羊成肌细胞激活素受体突变体
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