新型α-芋螺毒素基因克隆的引物筛选  

Primer Screening for New α-Conotoxin Gene Cloning

作　　者：郑晓冬[1,2] 高炳淼[1,2] 李宝珠[1,2] 彭超[1,2] 吴爱银 朱晓鹏[1,2] 陈心[1,2] 长孙东亭[1,2] 罗素兰[1,2] 

机构地区：[1]海南大学热带生物资源教育部重点实验室 [2]海南大学海口市海洋药物重点实验室,海南海口570228

出　　处：《生物技术》2011年第4期40-44,共5页Biotechnology

基　　金：重大新药创制国家科技重大专项课题(2009ZX09103-644);国家自然科学基金项目(30860368)资助

摘　　要：目的:优化PCR条件,建立能特异扩增出α-芋螺毒素基因片段的最理想PCR条件。方法:根据α-芋螺毒素基因保守的信号肽或内含子序列和非翻译区保守核苷酸序列设计了多组特异引物,并对引物浓度和退火温度等影响因素进行优化。结果:根据α-芋螺毒素基因保守的内含子序列为引物、引物浓度为0.1μmol/L、退火温度为50℃时,能特异的扩增出α-芋螺毒素基因片段,分子量大约分别为180bp和300bp。结论:采用优化的PCR条件,能筛选出克隆新型的α-芋螺毒素基因片段的最理想引物,为α-芋螺毒素的化学合成、活性研究和应用提供基础。Objective：To screen primers and optimize PCR conditions for more new α-conotoxin gene cloning.Method：According to the known conserved DNA sequence of signal or intron and the untranslated region（UTR） of α-conotoxin genes,many pairs of primers were designed,and also optimize the concentration of primer and the melting temperature.Result：With the known conserved DNA sequence of intron and primer 0.1μmol/L and the melting temperature of 50℃,α-conotoxin genes was obtained,which were 180bp and 300bp.Conclusion：The optimal primers were screened to obtain more new α-conotoxin genes from different cone snails＇ genomic DNA,which would be the basis for more novel α-conotoxins＇ discovery,chemical synthesis,bioassay and application.

关 键 词：α-芋螺毒素基因 基因组DNA 内含子 信号肽序列 保守性 PCR扩增 

分 类 号：Q523[生物学—生物化学]