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Cloning and vector construction of chicken ovalbumin gene regulatory sequences

  

  1. (Animal Husbandry and Veterinary Research Institute, Zhejiang Academy of Agricultural Sciences, Hangzhou 310021, China)
  • Online:2016-03-25 Published:2016-04-07

Abstract:  There was only one allele of ovalbumin gene in the chicken genome, but it could synthesize and secrete 2 g protein per day, which were accounting for more than 50% of the albumin protein and became the preferred choice in the regulation of exogenous gene expression. This study aimed to identify promoter enhancer and tissuespecific regional location factor by screening the optimization of ovalbumin gene promoter. The upstream -922~-2 073 and -2 801~-3 100 of ovalbumin promoter were divided into 12 regional which average sequence length were about 150 bp, and inserted into the upper reaches of -921~+38 sequences, those 12 series successfully constructed expression vectors provided materials for further optimization promoter with a shortened version. The first intron region of ovalbumin promoter was truncated around 300 bp of mini intron sequences, and successfully constructed 8 mini intron series of vectors. We also successfully separated chicken oviduct epithelial cells and optimized the electricity transfection conditions. In this study, the promoter region with strongest activity pGL4UP1412 and pGL4miniintron3 were screened through detected luciferase activity of the initial screening recombinant plasmid and intron recombinant, while inferred several regions containing enhancer sequence.

Key words: ovalbumin gene, regulation sequences, vector, luciferase activity