Scientia Agricultura Sinica ›› 2013, Vol. 46 ›› Issue (7): 1434-1440.doi: 10.3864/j.issn.0578-1752.2013.07.014

• ANIMAL SCIENCE·RESOURCE INSECT • Previous Articles     Next Articles

Construction of the Recombinant Adenovirus of FABP4 Gene of Qinchuan Cattle

 WEI  Sheng-Juan, ZAN  Lin-Sen, WANG  Hong-Bao, CHENG  Gong, JI  Shu-Han, WANG  Hong, FU  Chang-Zhen, JIANG  Bi-Jie   

  1. 1.College of Animal Science and Technology, Northwest A & F University, Yangling 712100, Shaanxi;
    2.National Beef Cattle Improvement Center in China, Yangling 712100, Shaanxi
  • Received:2012-10-16 Online:2013-04-01 Published:2013-01-30

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Abstract: 【Objective】The aim of this study was to construct the recombinant adenovirus vector with FABP4 gene of Chinese Qinchuan cattle, so as to provide a basis for studying FABP4 gene functions and mechanisms at cell level.【Method】The primers were designed according to the FABP4 mRNA sequence in GenBank, and the gene was cloned by RT-PCR and then sequenced. The bovine gene fragments containing both FABP4 gene and restriction enzyme were inserted into a shuttle vector pAdTrack-CMV to construct the recombinant shuttle vector pAdTrack-CMV-FABP4. After identifying by digestion and sequencing, pAdTrack-CMV-FABP4 plasmid was linearized by PmeⅠ, and then it was transformed into E. coli BJ5183 competent cells containing backbone vector pAdeasy-1 to obtain recombinant vector by homologous recombination. Then the positive plasmid was linearized by PacⅠ, and transfected into HEK 293A cells for virus packing, amplification and titer testing by TCID50.【Result】The results of enzyme digestion, sequencing and regular PCR detection showed that the recombinant overexpression vector containing FABP4 CDS region was successfully constructed. The infectious titer of the virus AD-FABP4 was 1.58×109 PFU•mL-1.【Conclusion】In this experiment, the recombinant adenovirus vector carrying FABP4 gene was constructed successfully and high titer adenovirus was acquired.

Key words: Qinchuan cattle , FABP4 gene , pAdTrack-CMV , pAdEasy-1 , recombinant adenovirus

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