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Abstract The target sequence in RSTN-T plasmid that contained pig resistin cDNA was amplified by PCR,inserted into eukaryotic expression vector pcDNA3.1(+), and then identified by restriction mapping and sequencing.The recombinant expression vector was transformed into Hela cell by LipofectaminTM 2000. The G418-resistant Hela cell clones were selected as well as the RSTN mRNA and the protein expression were analyzed by RT-PCR and Tricine-SDS-PAGE、Western blot,respectively. DNA sequencing demonstrated that pcDNA3.1-RSTN recombinant vector was constructed successfully. Hela cell clones were got by G418 selecting.The expression of pcDNA3.1-RSTN at mRNA level and protein level inHela cell was confirmed by RT-PCR and Tricine-SDS-PAGE、Western blot respectively. The successful construction and expression of pcDNA3.1-RSTN provides foundation for the further study of the pig resistin DNA vaccines.
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Received: 17 January 2007
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