利用重叠延伸PCR定点突变衰退病P23基因  

Overlap Extension PCR Based Site-Directed Mutants of CTV P23 Gene

作　　者：阳佳位[1,2] 王淼[2,3] 程春振[1,2] 魏召新[1,2] 陈凤娟[4] 

机构地区：[1]西南大学园艺园林学院,重庆400716 [2]西南大学柑桔研究所,重庆400712 [3]西南大学生命科学学院,重庆400716 [4]长江师范学院附属中学,重庆408003

出　　处：《西南大学学报（自然科学版）》2009年第12期43-47,共5页Journal of Southwest University(Natural Science Edition)

基　　金：国家自然科学基金资助项目(30571291);重庆市科委重大科技专项(CSTC;2007AA1018)

摘　　要：分析已知柑桔衰退病强毒系基因组并克隆CTV中一个重要的基因P23.利用重叠延伸PCR法的原理,成功的将P23基因保守序列第206位碱基由C突变成T,由此该位点成为限制性酶(SacI)的识别位点.把改良后的P23基因插入到T载体中,测序发现成功地突变掉目的位点.为进一步利用分子生物学手段研究CTV病毒,克隆突变CTV序列奠定了基础.Based on the analysis of CTV （citrus tristeza disease） genome,we cloned an essential gene P23.After the treatment of overlap extension PCR,the cytosine on the 206th base of the conserved sequence was mutated to thymine and the restriction enzyme site of SacI was mutated out.The modified gene P23 sequence was then inserted into the T vector.Sequencing showed that the targeted site was changed.The result is thought to have laid the foundation for cloning and mutation of CTV sequence with molecular biology tools for further analysis of CTV.

关 键 词：定点突变 重叠延伸PCR 柑桔 CTV病毒 

分 类 号：S666[农业科学—果树学] Q785[农业科学—园艺学]