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Construction of A Mammalian Expression Vector to Activate TumorAssociated AntigenSpecific T Lymphocytes

ObjectiveA mammalian expression vector encoding TAG72specific single chain variable region(scFv) fused to the transmembrane and intracellular domains of the signal transducing chain of CD28 was constructed, to generate for targeting of genetically modified T cells to gastrointestinal cancer, and to investigate the cytotoxicity against TAG72 positive target cells.MethodsThe transmembrane and intracellular domains of CD28 cDNA was amplified from human T lymphocytes using RTPCR to clone into a mammalin expression vecter, and CD28 fragments were ligated downstream of the antiTAG72 scFv cDNA and sequence verified.ResultsA 729base pair of antiTAG72 scFv was in accordance with sequence concerned; a 240base pair of cDNA of the transmembrane and intracellular domains of CD28 was confirmed as sequence concerning of Genebank.ConclusionWe constructed a mammalian expression vector encoding fusing gene to activate tumorassociated antigenspecific T lymphocyte, for generation of modified T lymphocytes to gastrointestinal tumors.