Construction and Identification of Eukaryotic Expression Recombinant Plasmid Containing Mice Connexin 43 Gene

Objective To construct eukaryotic expression recombinant plasmid containing mouse connexin 43 gene. Methods Total RNA was extracted from embryo mice. Connexin 43 cDNA was amplified by RT PCR, isolated and inserted into pBudCE4 1 eukaryotic expression plasmid. Then the recombinant plasmid was identified by enzyme digestion analysis and sequencing. Results The connexin 43 gene fragments of about 1 1 kb were specifically amplified by RT PCR technique. The recombinant plasmid digested by Hind Ⅲ and BamH Ⅰ was coincident with the expected connexin 43 cDNA fragment. The gene fragment containing of the whole coding sequence was successfully cloned in pBudCE4 1 plasmid. Conclusion Mouse recombinant plasmid pBudCE4 1/connexin 43 was successfully constructed.