畜牧兽医学报2011，Vol.42Issue(3)：343-348,6.
狮头鹅PRL基因的克隆及原核表达
Cloning and Prokaryotic Expression of PRL Gene in Shitou Goose
摘要
Abstract
To further study the physiological regulatory mechanisms of PRL on the reproductive performance of Shitou goose. The cDNA of goose prolactin (PRL) gene was amplified from anterior pituitary gland of Shitou goose by RT-PCR. Then the PCR product was cloned into the pMD18-T vector to construct the pMD18-PRL for sequencing. And then the cDNA was subcloned into the prokaryotic expressing vector pET32a (+). Subsequently, the recombinant plasmid pET32-PRL was transformed into the E. coli BL21 (DE3) expression bacteria, then the recombinant PRL was induced by IPTG. The purified recombinant PRL was detected by SDSPAGE and Western Blot. The results showed that coding region of PRL gene was comprised of 600 nucleotides(GenBank No. GQ856665) and encoded 199 amino acids putative protein which shared highly conservation with that of other birds. It was found that PRL protein was comprised of several α-Helixes, β-Sheets and Coils. It was inferred that the superiority region of antigenic epitope were in the N' 70-76,95-102,150-155 and 207-213 sections. A high proportion of soluble PRL fusion protein about 53. 6％ of total bacterial protein was obtained in E. coli BL21 (DE3).The molecular weight of the fusion protein was about 41 ku. The Western Blot result showed that the recombinant protein was recognized by antiserum specifically. The result of this study will provide basis for further study of the biological function of prolactin protein and the effects of PRL on the broodiness and production traits of Shitou goose.关键词
狮头鹅/催乳素/克隆/基因表达PRL蛋白分类
农业科技引用本文复制引用
贾汝敏,吴慧英,刘铀,张丽..狮头鹅PRL基因的克隆及原核表达[J].畜牧兽医学报,2011,42(3):343-348,6.基金项目
湛江市科技招标项目(0609135) （0609135）
广东海洋大学基金项目(C04095) （C04095）
校企合作项目(0810314) （0810314）
