论文部分内容阅读
为构建猪RBP?4基因原核表达载体，并在大肠杆菌中表达蛋白。取成年母猪正常卵巢组织提取总RNA，RT－PCR后回收扩增产物，构建表达载体pEASY－E1－RBP4，转化BL21（ DE3） pLysS，IPTG诱导表达。测序结果显示，RT－PCR获得的cDNA全长序列与GenBank中的序列基本一致。原核表达产物经SDS－PAGE电泳鉴定，其以包涵体形式表达。试验成功构建了猪RBP4基因原核表达载体，并在大肠杆菌中表达，为后续蛋白纯化及抗体的制备奠定基础。“,”The objective of this study is to construct prokaryotic expression vector of swine RBP?4 gene and express it in E.coli. The total RNA was extracted from swine normal ovarian tissue. The se?quence including the whole length of RBP?4 was amplified by RT-PCR and inserted into pEASY?E1, and then transformed into E.coli BL21(DE3)pLysS and inducted by IPTG. The sequencing re?sult proved that the cloned RBP?4 cDNA sequence was uniform with its sequence stored in GenBank. The expression product of RBP?4 gene was identified by SDS-PAGE and it was expressed in the form of inclusion bodies. Prokaryotic expression vector of RBP?4 gene was successfully established, and the gene was expressed in E.coli, which was prepared for purification and RBP?4 polyclone anti?body.