Construction and identification of the RNA interference eukaryotic expression vector targeting the CD40 gene in rats / 医学研究生学报

ABSTRACT

Objective: To construct and identify the eukaryotic expression vector targeting the CD40 gene in rats.Methods: Two sequences corresponding to the rat CD40 gene were designed on Ambion's Web.The two complementary oligonucleotide strands of DNA fragments were synthesized by chemosynthesis.After annealing of the complementary strands,the DNA fragments were connected to the polyclone sites of the pSilencer 4.1-CMV neo vector,followed by transformation,amplification,plasmid extraction and identification of the recombinant plasmids by BamH Ⅰ and Hind Ⅲ digestion and DNA sequence analysis.Results: The connections between the DNA fragments encoding CD40-targeted siRNA and the pSilencer 4.1-CMV neo vector were correct,as confirmed by agarose gel electrophoresis and DNA sequence analysis.Conclusion: The two recombinant plasmids expressing siRNA of the rat CD40 gene were successfully constructed,which prepared the groundwork for future research on the role of the CD40 gene in homograft rejection.