桂北五步蛇金属蛋白酶原基因1137bp片段的克隆和序列分析

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pGEM-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：RTPCR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91％的同源性。

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

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%.