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Progress in Modern Biomedicine. 2014, (04)

Construction of Lentiviral Vector Carrying ZO-1 GeneChinese Full Text

SHENG Tao;ZENG Jian-ping;LI Hui;Department of Cardiology,The Second Xiangya Hospital of Central South University;Department of Cardiology,Xiangtan Central Hospital;Reproductive center,Xiangtan Central Hospital;

Abstract: Objective: The joint of adjacent heart muscle fibers is called myo cardial intercalated disc, which is important for myocardial electrical activity. It has been widely concerned about the study of intercalated disc related gene Cx43. Under different pathological status, it is related to another gene ZO-1. However, the research of ZO-1 is scanty, so we cloned the ZO-1 gene of porcine into the lentiviral vector and identificate the recombinant plasmid,which will lay the foundation for further research. Methods: The porcine ZO-1 gene was amplified by PCR, and cloned into lentiviral expression vector. The reconstructed Lenti- EF1α-EGFP-TRE-ZO-1 plasmid was identified by transformation, plasmid extraction, double enzyme digestion and DNA sequencing analysis. Results: The porcine ZO-1 DNA sequences were successfully inserted into the lentiviral vector.The reconstructed plasmid was identified correctly by PCR and double enzyme digestion.The recombinant Lenti-EF1α-EGFP-TRE-ZO-1 vector was confirmed by DNA sequencing. Conclusion: Lenti-EF1α-EGFP-TRE-ZO-1 lentiviral vector has been successfully constructed, so we can use it to infect related cells or animals, which will be useful for further study in vitro and in vivo, to understand its function in different pathological status.
Keywords：

ZO-1; Gene cloning; Lentiviral vector;

  • DOI：

    10.13241/j.cnki.pmb.2014.04.007

  • Series：

  • Subject：

  • Classification Code：

    R346

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