Local Transfection Of The C-type Natriuretic Peptide Genes Control Vascular Anastomotic Restenosis Study

Objective To clone human C-type natriuretic peptide gene and reconstruct its eukaryotic expressive vector for future investigation of gene therapy.Methods The full coding domain sequence of human C-type natriuretic peptide gene was cloned from human placenta with RT-PCR and was sub-cloned into pBS-T. After screening of negative pBS-T/CNP with restriction enzyme Smaâ… degestion, the targeted DNA fragment, digested with Hindâ…¢and BamHâ… , was directional cloned into eukaryotic expressive plasmid pcDNA3.1(+). Then, the reconstructed plasmid was identified with enzyme digestion, PCR and sequencing.Results It was demonstrated that the full coding domain sequence of human C-type natriuretic peptide gene was accurately cloned into digestion sites between Hindâ…¢and BamHâ… in pcDNA3.1(+) without mutation and transposition.Conclusion The successive reconstruction and verifying of eukaryotic...