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Cloning and Eukaryotic Expression of Adenovirus Type 7 E1A Gene
Xin-Feng Shen, Bao-Ling Shen, Yan-Yan Huang, Xue-Jiao Yu, Ying Peng*, Jian-Xin Lu*
Zhejiang Provincial Key Laboratory of Medical Genetics, Wenzhou Medical College, Wenzhou 325035, China)
Abstract: To provided data and material for the future study on the pathogenic mechanism of the adenovirus type 7 (Ad7) E1A， we isolated the Ad7d2 and constructed eukaryotic expression vector of Ad7 E1A gene. Respiratory specimens were inoculated into A549 cells to isolate adenovirus and extract viral DNA. Overlap extension PCR was applied to amplify the exon of E1A gene. PCR products were cloned into the pIRES-Neo vector to produce the expression vector pIRES-Neo-Ad7E1A. After transfection to A549 cell lines， Western blot was applied to identify the expression products. The correct clone of complete cDNA of Ad7 E1A were confirmed by DNA sequencing. The recombinant E1A products were identified by Western blot. The eukaryotic expression system of adenovirus type 7 E1A gene was constructed.
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