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牛肌肉生成抑制素基因功能区的克隆与原核表达
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吕文发，袁天祥，孔振兴
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（吉林农业大学 动物科技学院，吉林 长春130118）
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摘要：
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根据GenBank中牛肌肉生成抑制素（MSTN）基因序列设计了1对引物，在引物两端分别加EcoRⅠ和XhoⅠ识别位点。利用RT-PCR技术扩增出了牛MSTN功能区序列。分别构建克隆和原核表达载体，酶切、PCR鉴定及测序分析表明,该基因功能区序列的克隆载体和原核表达载体已成功构建。筛选阳性菌，经IPTG诱导,牛MSTN基因功能区在大肠埃希氏菌中成功表达。 |
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关键词：
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肌肉生成抑制素基因；牛；RT-PCR；原核表达 |
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中图分类号：S 818.9：Q 786文献标识码：
A 文章编号：1673-4696(2006)06-0482-03 |
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Cloning and prokaryotic expression of bovine myostatin gene |
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LU Wen-fa， YUAN Tian-xiang，KONG Zhen-xing |
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(College of Animal Science and Technology, Jilin Agricultural University, Changchun 130118, China) |
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Abstract: In order to express the functional sequence of bovine myostatin gene in the prokaryotic system, a pair of primers was designed according to the myostatin cDNA sequence and optimized with restriction sites for two restriction endonucleases EcoRⅠ and XhoⅠ. The functional fragment was amplified from bovine total RNA using RT-PCR. The cloning plasmid and pET-28a vector were digested, retrieved and sequenced, respectively. Double digestion with EcoRⅠ and XhoⅠ, PCR identification and sequence analysis showed that both the cloning vector and the prokaryotic expression vector for the functional fragment of the myostatin gene were constructed successfully. Positive clones were selected and induced by IPTG, and the expressed product of the bovine functional fragment was obtained. |
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Key words: myostatin gene; cattle; RT-PCR; prokaryotic expression |
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