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一种用高拷贝质粒载体制备BAC载体基本功能基因的新方法(英文)

Research on the Construction of Bacterial Artificial Chromosome Vector DNA and the Potentials in Application

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【作者】 崔红玉王亚萍徐明举薛永志石星明兰德松王云峰童光志

【Author】 CUI Hong-yu1,WANG Ya-ping2,XU Ming-ju1,XUE Yong-zhi1,SHI Xing-ming3,LAN De-song3,WANG Yun-feng3,TONG Guang-zhi31. Department of Animal Medicine,Hebei North University,Zhangjiakou 075131; 2. Animal Quarantine Station of Xuanhua District,Zhangjiakou City,Hebei Province,Zhangjiakou 075131; 3. National Key Laboratory of Veterinary Biotechnology,Harbin Veterinary Research Institute,Chinese Academy of Agricultural Sciences,Harbin 150001

【机构】 河北北方学院动物科技学院动物医学系河北省张家口市宣化区动物检疫站中国农业科学院哈尔滨兽医研究所兽医生物技术国家重点实验室

【摘要】 [Objective] The aim of this study was to provide a method for solving the problems in preparing BAC vector with High-copy plasmid pUC119-Bluelox BAC.[Method] With selecting a proper single restriction site,sequences of a single copy BAC vector plasmid were inserted into proper site of High-copy plasmid pUC119 vector.[Result] The gene sequence of BAC vector lost control function of single copy number in new plasmid pUC119-BAC and was copied through High-copy form. The gene sequence of BAC vector basic function was completely cutted off through single enzyme digestion and the control function of single copy could be recovered by auto-connection.[Conclusion] The High-copy pUC119-BAC plasmid was used to copy and amplify high copy of basic function gene sequence in BAC vector,besides that it could be used to construct transfer vector of molecular cloned recombinant virus or BAC library.

【Abstract】 [Objective] The aim of this study was to provide a method for solving the problems in preparing BAC vector with High-copy plasmid pUC119-Bluelox BAC.[Method] With selecting a proper single restriction site,sequences of a single copy BAC vector plasmid were inserted into proper site of High-copy plasmid pUC119 vector.[Result] The gene sequence of BAC vector lost control function of single copy number in new plasmid pUC119-BAC and was copied through High-copy form. The gene sequence of BAC vector basic function was completely cutted off through single enzyme digestion and the control function of single copy could be recovered by auto-connection.[Conclusion] The High-copy pUC119-BAC plasmid was used to copy and amplify high copy of basic function gene sequence in BAC vector,besides that it could be used to construct transfer vector of molecular cloned recombinant virus or BAC library.

  • 【文献出处】 Agricultural Science & Technology ,农业科学与技术(英文版) , 编辑部邮箱 ,2008年02期
  • 【分类号】S188
  • 【下载频次】101
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