海南山蛭蛭素基因的克隆和序列分析  

Cloning and Sequencing of Haemadin Gene from Haemadipsa hainana

作　　者：谭琳[1] 康由发[2] 谭恩光[3] 郑学勤[2] 

机构地区：[1]海南医学院生化教研室,海口市571101 [2]中国热带农业科学院热带作物生物技术国家重点实验室,海口市571101 [3]中山医科大学生物教研室,广州市510089

出　　处：《热带农业科学》2004年第2期23-25,61,共4页Chinese Journal of Tropical Agriculture

摘　　要：以海南山蛭(Haemadipsahainana)基因组DNA为模板,以合成的2段30个寡聚核苷酸的序列为引物,经过PCR扩增,分离海南山蛭中的蛭素基因,获得了1条大小约237 bp的DNA片段。将其克隆到pGEM-Teasy载体中,经筛选与检测,并进行序列分析。结果显示,所克隆的海南山蛭蛭素基因的大小为231 bp,与Dr. Burkhard报道的印度山蛭蛭素cDNA序列相比,其核苷酸的同源性为99.9%。A specific DNA fragment about 237 bp was obtained from PCR in which the genomic DNA from Haemadipsa hainana served as template and two short specific sequences of DNA as primers. By electro phoresis on low melting agrose gel, the specific DNA fragment was retrieved, purified and then linked into pGEM-Teasy vector by linkase T4DNA. The recombinant plasmid was transfered into Escherichia coli DH5α. The transformer was first screened by IPTG/X-gal, then authenticated by PCR and by SacⅠ and ApaⅠ restriction enzyme digestion. The bacteria containing the recombinant plasmid were quickly proliferated, and the heterogeneous gene was magnified. Purified recombinant plasmid was sequenced. The result showed that the sequence of haemadin gene had a 99.9% homology with cDNA sequence of Haemadin from an Indian leech.

关 键 词：海南 山蛭 蛭素基因 基因克隆 序列分析 

分 类 号：S899[农业科学—特种经济动物饲养]