东北地方猪抑肌素基因表达载体的构建及其原核表达的研究

瘦肉率的提高是猪育种中重要的目标性状之一,其大小与骨骼肌的量密切相关.1997年由McPherror等发现的肌肉生长抑制素基因(myostatin,简写为MSTN)因其功能丧失后会导致小鼠骨骼肌的量极显著增加而引起广泛关注. 为进一步研究myostatin基因的本质功能,奠定猪遗传改良和分子育种基础,本研究以克隆在pGEM-3Z上的猪myostatin基因cDNA为研究对象,利用Oligo 5.0软件设计出两对上、下游特异性引物,并分别引入EcoRI和BamHI两酶切位点.扩增出与表达载体相匹配的全长与半长MSTN基因cDNA序列,构建融合表达载体,进而将构建好的表达载体转化到大肠杆菌BL-21中表达出全长与半长蛋白,实现了具有二硫键基因在大肠杆菌中的正确表达.

CONSTRUCTING PORCINE MYOSTATION GENE EXPRESSING VECTOR AND EXPRESSING IT IN E. COLI

Lean meat percentage is one of the most important economic traits in pig breeding programs.Myostatin was identify by Mcpherron(1997)as negative regulator of muscle mass in mice. In order to study further natural functions of the porcine myostatin gene of which were obtain PGEM-3Z cloning vector in myostatin gene and to establish theory basis of molecular breeding of porcine,two pairs of upper,lower specific,PCR primers which containing EcoR I and BamH I restriction enzyme sites was designed.The full-length and half-length myostatin cDNA was amplified to construct fusion-expressing vector and to transform E.coli BL-21 cell expressing full and half length protein.We have achieved accurate expression taken on disulfide bond (linage) gene.