Title: Cloning and Sequence Analysis of Porcine Interferon Gamma Gene and Construction of Recombinant Adenovirus Vector Containing IFN-γ
Abstract: The total RNA was isolated from spleen lymphocytes from swine,then the IFN-γ gene was amplified via RT-PCR.The gene segment of IFN-γ was cloned into the vector pGEM-T.From the sequence,it was found that the IFN-γ gene was consisted of 517 nucleotides.The ORF was consisted of 501 nucleotides,encoding 166 amino acids.The homology of the IFN-γ gene was 100% at nucleotide level and amino level compared with other porcine IFN-γ sequence published on GenBank.The objective gene was cloned into pAd-Shuttle-CMV vector and constructed the eukaryotic expression plasmid pAd-Shuttle-CMV-IFN-γ.The positive recombinant pAd-Shuttle-CMV-IFN-γ was linearized and transformed into Ad-BJ5183 competent cells which had already been transformed with adenovirus skeletal vector pAdeasy-1.The Adenovirus expression plasmid was confirmed by Pac Ⅰ digestion.These results indicated that the recombinant adenovirus plasmid was successfully constructed and established basic work for further study.
Publication Year: 2006
Publication Date: 2006-01-01
Language: en
Type: article
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