牛蛙(Rana catesbeiana)蛙皮抗菌肽基因的克隆、测序及其表达  

Cloning and expression of antimicrobial peptides gene from bullfrog skin

作　　者：韩俊友[1] 乔红伟[2] 赵瑞利[1] 胡博[1] 谢芳[1] 王金玲[3] 郑伟[4] 雷连成[1] 韩文瑜[1] 

机构地区：[1]吉林大学畜牧兽医学院,吉林长春130062 [2]中国医学科学院中国协和医科大学实验动物研究所,北京100021 [3]沈阳出入境检验检疫局,辽宁沈阳110016 [4]吉林省水产科学研究院,吉林长春130033

出　　处：《中国兽医学报》2007年第3期399-402,共4页Chinese Journal of Veterinary Science

基　　金：国家自然科学基金资助项目(30571416)

摘　　要：利用RT-PCR的方法从牛蛙皮肤组织中克隆到大小为270bp的片段RCABP,将其克隆到pGEM-T载体,测序获得1个新的碱基序列。将此基因克隆到原核表达载体pQE-80L,获得融合表达质粒pQE-80L/DHFR/ABP,在1%IPTG诱导下进行表达。SDS-PAGE检测表明,重组蛙皮抗菌肽蛋白的表达量占菌体总蛋白的24%,以包涵体形式存在。体外抑菌试验表明,所构建的质粒能在大肠杆菌中表达具有体外抑菌活性的蛙皮抗菌肽,该融合蛋白具有良好的应用前景。Using RT-PCR techuique, a nucleotide segment of 270 bp was obtained from the skin of bullfrog and then cloned into the vevtor of pPGEM-T. The product was gained and sequenced. The identified cDNA was cloned into the new type of prokaryotic expression vector pQE-80L, and a synthetized expression plasmid named pQE- 80L/DHFR/ABP obtained. The E. coli competent host BL21（DE3）lysS was transformed with pQE-80L/DHFR/ ABP and the antimicrobial peptide gene was expressed at a high level, amounting for 24% of the induced recombi- nant bacteria protein at the presence of IPTG.

关 键 词：牛蛙 抗茵肽 基因表达 

分 类 号：Q78[生物学—分子生物学] S852.4[农业科学—基础兽医学]