桂北五步蛇金属蛋白酶原基因1137bp片段的克隆和序列分析  

STUDY OF THE METALLOPROTEINASE GENES OF AGKISTRODON ACUTUS VENOM FROM NORTHERN MOUNTAIN AREA OF GUANGXI

作　　者：杨海波[1] 樊晓晖[1] 侯小琼[1] 黄企光[1] 黎肇炎[1] 

机构地区：[1]广西医科大学微生物与免疫学教研室,南宁530021

出　　处：《广西医科大学学报》2004年第6期824-827,共4页Journal of Guangxi Medical University

基　　金：广西青年科学基金资助项目 ( No.桂青科 9912 0 18)

摘　　要：目的 :克隆桂北五步蛇金属蛋白酶原基因 ,并对其进行序列分析。方法 :采用一步法抽提广西桂北山区五步蛇蛇毒总RNA,经 RT- PCR扩增纤溶酶金属蛋白酶原基因 ,将扩增产物克隆至 p GEM- T Easy载体 ,挑选白色菌落 ,用酶切和 PCR法对其进行鉴定。直接利用纯化 PCR产物或提取阳性菌落进行测序并推导其编码的氨基酸序列。结果 :RT- PCR和 PCR扩增得到一约 1137bp产物 ,并将其克隆及测序。结论 :和已报道的皖南五步蛇纤溶酶金属蛋白酶氨基酸序列相比较 ,二者之间有Objective:To clone and sequence a cDNA encoding fibrinolysin metalloproteinase from the venom of Agkistrodon acutus from Guangxi.Methods:One step method was used to extract total RNA from the venom of Agkistrodon acutus found in northern mountain area of Guangxi. The DNAs encoding fibrinolysin metalloproteinase were amplified by one step method (RT-PCR and PCR reactions occurred in the same tube). The 1 137 bp PCR product was cloned into the pGEM-T Easy vector. Plasmids obtained from positive clones were identified by means of digestion with EcoR I and PCR reaction. The 1 137 bp PCR product was sequenced. Result:We got 1 137 bp amplified product that was cloned into E.coli JM109. Its sequence was determined.Conclusion:Compared with the cDNA sequence for the fibrinolysin metalloproteinase from the venom of Agkistrodon acutus from the south of Anhui Province, their homology is 91%.

关 键 词：五步蛇 金属蛋白酶 序列分析 

分 类 号：Q959.6[生物学—动物学] Q55