Construction of a recombinant adenovirus vector of the rat Lys184del mutant cardiac troponin I gene / 医学研究生学报

ABSTRACT

Objective: To construct a recombinant adenovirus expression vector of the SD rat Lys184del mutant cardiac troponin I gene(cTnI Lys184del).Methods: The Lys184del mutant cardiac troponin I gene of SD rat was induced by PCR-based site-directed mutagenesis,the amplified fragment containing the mutation was then inserted into the cloning vector pMD18-T.After the addition of a sequence encoding a 6?His tag at the C-terminus by PCR,the coding region of cTnI Lys184del was subcloned into Adeno-X viral DNA to form a recombinant adenoviral plasmid,which was packed into infectious recombinant adenoviral particles(Ad-cTnI Lys184del) by transfecting HEK293 cells.Results: The DNA sequence of Ad-cTnI Lys184del was proved to be correct by PCR and sequencing,and its expression was identified by Western blot analysis.Conclusion: The successfully constructed recombinant adenovirus Ad-cTnI Lys184del has provided valuable data for further researches on its functional characteristics.