肉牛肌生成抑制素成熟蛋白编码序列的克隆与原核表达

Cloning and Prokaryotic Expression of the Coding Sequence of Maturation Protein of Bovine Myostatin Gene

[目的]原核表达肉牛肌生成抑制素成熟蛋白编码序列基因并进行功能验证,为后期的小鼠接种实验打下基础。[方法]采用RT-PCR方法扩增肉牛MSTN成熟蛋白编码序列基因,该扩增产物经过双酶切后克隆到表达载体pET28a(+)中,转化大肠杆菌BL21,经IPTG诱导表达后采用SDS-PAGE和Western-blot检测重组目的蛋白的表达结果。[结果]经Anti-His-Tag鉴定正确。[结论]肉牛MSTN成熟蛋白编码序列基因在原核表达系统中得到正确表达。

[Objective] The aim was to test and verify maturation protein of bovine myostatin（MSTN） gene based on prokaryotic expression,to provide basis for inoculation experiment of mice.[Method] RT-PCR method was used to amplify coding sequence of maturation protein of bovine myostatin（MSTN） gene,then after double digestion,amplification products were cloned to expression vector pET28a（＋）,transform escherichia coli BL21,after induction of IPTG,SDS-PAGE and Western-blot were used to detect expression of results of recombinant protein.[Result] Anti-His-Tag identification was correct.[Conclusion] Beef cattle MSTN were expressed in prokaryotic expression system correctly.