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紫杉醇酰化酶多基因家族成员克隆时的引物设计与PCR参数优化 被引量：1

Primer Design and PCR-Parameter Optimization for Cloning the Genes of Taxol Acyltansferase Superfamily
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摘要 紫杉醇酰化酶多基因家族是催化红豆杉紫杉醇合成过程中的各酰化酶基因的总和.该基因家族的不同成员之间具有很高的保守性,本实验以该基因家族中的紫杉烷2α-O-苯甲酰转运酶(TBT)基因克隆为例,从引物设计与PCR参数优化角度,总结了针对类似高保守性多基因家族的基因克隆的经验,表明分别在保守性极高部位设计第一次扩增引物及在保守性极低部位设计巢式引物,并采用touchdow n PCR程序及高效能的T aq DNA聚合酶是成功的关键. Taxol acyltansferase superfamily is the general term of various acyltansferase genes catalyzing taxol biosynthesis in Taxus and its members are highly conservative among themselves. Using taxane 2α-O-benzoyltransferase gene cloning as an example,this study summarized the cloning experiences of the genes of similar and highly conservative super families and indicated that the keys of successful primer design were to design the primers of the first amplification in highly conservative regions and nested primers in lowly conservative regions as well as to adopt touchdown PCR protocol and Taq DNA polymerase.
作者 赵东利 逯越 黄胜英 李雅杰
机构地区 大连大学医学院
出处 《西北植物学报》 CAS CSCD 北大核心 2005年第11期2178-2182,共5页 Acta Botanica Boreali-Occidentalia Sinica
关键词 多基因家族 紫杉醇酰化酶 引物设计 基因克隆 红豆杉 superfamily taxol acyltansferase primer design gene cloning Taxus
分类号 Q785 [生物学—分子生物学]
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参考文献6

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同被引文献17

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西北植物学报
2005年 第11期

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