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两种pUC18高效T载体的构建

Construction of Two Highly Efficient pUC18 T-vectors

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【作者】 刘勇王立良袁力勇陈尔佳庄俊英孙茂盛戴长柏

【Author】 LIU Yong, WANG Li liang 1) , YUAN Li yong,CHEN Er jia, ZHUANG Jun ying,SUN Mao sheng,DAI Chang bai (Institute of Medical Biology,CAMS and PUMC,Kunming 650118,China; 1) Institute of Basic Medical Sciences,CAMS and PUMC,Beijing 10000

【机构】 中国医学科学院中国协和医科大学医学生物学研究所!昆明6501181,中国医学科学院中国协和医科大学基础医学研究所!北京100005,中国医学科学院中国协和医科大学医学生物学研究所!昆明6501181,中国医学科学院中国协和医科大学医学生物学研究所!昆明6501181,中国医学科学院中国?

【Abstract】 Two T vectors were generated by restriction endonuclease Xcm Ⅰ instead of Taq or other DNA polymerases to create the 3’T over hangs.A fragment of adenoviral genome position 10659-11865 was amplified by PCR and Xcm Ⅰ recognition sites were introduced to both ends of the PCR product.This fragment was cloned into the Sma Ⅰ site of pUC18 or the linearized pUC18 with its polylinker deleted.The recombinant plamids were cleaved with Xcm Ⅰ.the larger fragments generated which had 3’T over hangs at both ends were used as T vctors.Genes of rotavirus VP7 and the plasminogen k5 were successfully cloned into these two T vectors with recombination efficiency (recombinants/transformants×100%) of 100%,no blue/white clolny screening assay was needed.

  • 【文献出处】 中国生物化学与分子生物学报 ,CHINESE JOURNAL OF BIOCHEMISTRY AND MOLECULAR BIOLOGY , 编辑部邮箱 ,2000年04期
  • 【分类号】Q784
  • 【被引频次】5
  • 【下载频次】433
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