生物技术通报2016，Vol.32Issue(5)：82-90,9.DOI:10.13560/j.cnki.biotech.bull.1985.2016.05.011
新疆南部和田羊、卡拉库尔羊、多浪羊MSTN基因的真核表达
Eukaryotic Expression of MSTN Gene from Hetian Sheep,Carla Kul Sheep,and Duolang Sheep in the Southern Xinjiang
摘要
Abstract
This work aims to construct the fusion expression plasmids ofmyostatin gene(MSTN)from 4 local varieties(lines)sheep, namely plain Hetian sheep(HP),mountain Hetian sheep(HS),Carla Kul sheep(KL),and Duolang sheep(DL)in the southern Xinjiang with enhanced green fluorescent protein(EGFP),and the expression ofMSTN in eukaryotic cells. Total RNA was extracted from the skeletal muscle in 4 varieties of sheep,cDNA was acquired by reverse transcription,andMSTNamplified by PCR was ligated with pMD18-T vector to construct recombinant plasmid and sequenced. ThenMSTN(HP,HS,DL,and KL)gene was inserted into the fusion expression vector pEGFP-N1,the recombinant plasmid of pEGFP-N1-MSTNin aeukaryotic fusion expression was constructed. The plasmid with over 80% cells covered was transfected into CHO by liposome,the green fluorescent of fusion protein of EGFP and MSTN in CHO cells was observed. The results showed that the ORF sequence ofMSTN gene in any of HP,HS,KL,and DL was 1 125 bp(excluding stop codon),encoding 375 amino acids. Comparing MSTN sequences in GenBank,there was differences in HS and HP;and 2 nucleotide variation in KL caused 2 amino acid mutations;and 6 nucleotide differences in DL resulted in 4 amino acid variations. In conclusion,the recombinant plasmid for EFGP and MSTN co-expression was constructed and expressed in eukaryotic CHO cells.关键词
和田羊/卡拉库尔羊/多浪羊/肌生成抑制素/CHO细胞/绿色荧光蛋白Key words
hetian sheep/carla kul sheep/duolang sheep/myostatin/CHO cell/green fluorescent protein引用本文复制引用
王海涛,金波,李树伟..新疆南部和田羊、卡拉库尔羊、多浪羊MSTN基因的真核表达[J].生物技术通报,2016,32(5):82-90,9.基金项目
新疆生产建设兵团博士资金专项 （）
