Objective : to clone and sequence the cdna encoding metalloproteinase from the venom of agkistrodon acutus from northen mountain area of guangxi province . methods : one step method was used to extract total rna from the venom of agkistrodon acutus found in northern mountain area of guangxi province . different kinds of cdna encoding metalloproteinase were amplified by one step method ( rt - pcr and pcr reactions occurred in the same tube ) using different primers 方法：从桂北五步蛇毒腺中抽提总rna ，利用不同的引物，采用一步法（ rt - pcr和pcr在同一管内进行）扩增出不同的dna条带，利用平端连接的方法将pcr扩增产物克隆至pgem - teasy载体，转化大肠杆菌jm109 ，挑选白色菌落提取质粒，用pcr对其进行鉴定，直接利用纯化pcr产物或提取阳性菌落质粒进行测序。
