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2007•Journal of Jiangsu UniversityRequires access

Molecular cloning and sequence analysis of human Glucocorticoid-inducible tumor necrosis factor receptor gene

Min Yang

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Abstract

Objective: To clone and analyze a full-length cDNA encoding human glucocorticoid-inducible tumor necrosis factor receptor(GITR) gene.Methods: The cDNA of GITR was amplified by RT-PCR using the total RNA extracted from normal human peripheral blood mononuclear cell(PBMC).The PCR product was inserted into pGEM-T vector and then transformed into E.coli DH5α.The positive recombinant clone was analyzed by digestion of restriction endonuclease and DNA sequencing. Results: The cDNA of human GITR had a complete open reading frame with a length of 726 bp,which encoded a product of 241 amino acid,and shared 100% homology with the sequence of mRNA for GITR in Genbank. Conclusion: The cDNA of hGITR was cloned successfully,which posed a basis for further researching on its biological function.

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What this paper is about

Objective: To clone and analyze a full-length cDNA encoding human glucocorticoid-inducible tumor necrosis factor receptor(GITR) gene.Methods: The cDNA of GITR was amplified by RT-PCR using the total RNA extracted from normal human peripheral blood mononuclear cell(PBMC).The PCR product was inserted into pGEM-T vector and then transformed into E.coli DH5α.The positive recombinant clone was analyzed by digestion of restriction endonuclease and DNA sequencing. Results: The cDNA of human GITR had a complete open reading frame with a length of 726 bp,which encoded a product of 241 amino acid,and shared 100% homology with the sequence of mRNA for GITR in Genbank. Conclusion: The cDNA of hGITR was cloned successfully,which posed a basis for further researching on its biological function.

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Available abstract

Objective: To clone and analyze a full-length cDNA encoding human glucocorticoid-inducible tumor necrosis factor receptor(GITR) gene.Methods: The cDNA of GITR was amplified by RT-PCR using the total RNA extracted from normal human peripheral blood mononuclear cell(PBMC).The PCR product was inserted into pGEM-T vector and then transformed into E.coli DH5α.The positive recombinant clone was analyzed by digestion of restriction endonuclease and DNA sequencing. Results: The cDNA of human GITR had a complete open reading frame with a length of 726 bp,which encoded a product of 241 amino acid,and shared 100% homology with the sequence of mRNA for GITR in Genbank. Conclusion: The cDNA of hGITR was cloned successfully,which posed a basis for further researching on its biological function.

Key concepts: Complementary DNA, Biology, Molecular biology, Gene, Restriction enzyme, Recombinant DNA, Open reading frame, Molecular cloning

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Molecular cloning and sequence analysis of human Glucocorticoid-inducible tumor necrosis factor receptor gene — Research Paper | ScholarLens