猪Ghrelin基因的克隆及原核表达  

Chemical Synthesis and Prokaryotic Expression of Ghrelin of Pig

作　　者：杨连玉[1] 杨文艳[1] 赵颖彩[2] 钱剑[3] 王哲[1] 

机构地区：[1]吉林大学畜牧兽医学院,吉林长春130062 [2]吉林农业大学动物科技学院,吉林长春130118 [3]塔里木农垦大学动物科技学院,新疆阿克苏830000

出　　处：《中国兽医学报》2005年第6期614-616,共3页Chinese Journal of Veterinary Science

基　　金：国家自然科学基金资助项目(30170713);吉林省教育厅资助项目(200401)

摘　　要：从猪下丘脑、胃等组织中提取总RNA,根据已发表的猪的G hre lin mRNA序列设计合成引物,通过RT-PCR进行cDNA扩增,获得了282 bp的片段。将该片段克隆于pM D-18T载体后进行序列分析,确认PCR产物为G hre lin cD-NA。从阳性克隆中提取质粒,经N heⅠ和X hoⅠ双酶切,回收282 bp的目的片段,定向克隆到pET-28a表达载体中,提取质粒并再次转化到BL 21(DE 3)中,成功地筛选出阳性克隆。经IPTG诱导阳性菌,通过SDS-PAGE检测出猪G hre lin基因的表达。Fourty eight strains were isolated from diseased pigs in Qinhuangdao and Tangshan of Hebei province from 2003 to 2004. The morphological,cultural and biochemistry characters indicated that the isolates belonged to genus Streptococcus. Furthermore,43 isolates could be agglutinated by the Lancefield group A-G latex diagnostic kit. The positive samples were 23, 18,1 and 1 strains for group D,C,B and A respectively. But the rest five serums could not be typied. The isolates were demonstrated to be pathogenicity to mouse,rabbit and pig. The results showed that swine Strep. suis were mainly caused by groups D in these area.

关 键 词：GHRELIN 基因克隆 原核表达 猪 

分 类 号：Q78[生物学—分子生物学]