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2005•Journal of Jiangsu UniversityRequires access

Cloning of Specific Transcription Factor T-bet of Murine T Helper 1 Type Cells

Wang Suo-ying, XU Hua-xi

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Abstract

Objective: To clone all-length coding cDNA of specific transcription factor T-bet of murine T helper 1 type cells.Methods: Total RNA as well as mRNA were isolated from splenic cells of Balb/c mice and the entire coding cDNA sequence of T-bet was amplified by reverse transcription polymerase chain reaction(RT-PCR).The T-bet cDNA were cloned into the pGEM-T vector with a T base at 3' terminal.The identification was made by means of restriction enzyme analysis,PCR and DNA sequencing.Results: The murine entire coding cDNA of 1592bp have been amplified from spleen cells.There was 99.8% homogenous nucleotide sequence as compared with standard sequence of T-bet from Genbank.Conclusion: We successfully cloned the cDNA of murine specific transcription factor T-bet.It will be the basis for studying immune interference of transferring gene.

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Objective: To clone all-length coding cDNA of specific transcription factor T-bet of murine T helper 1 type cells.Methods: Total RNA as well as mRNA were isolated from splenic cells of Balb/c mice and the entire coding cDNA sequence of T-bet was amplified by reverse transcription polymerase chain reaction(RT-PCR).The T-bet cDNA were cloned into the pGEM-T vector with a T base at 3' terminal.The identification was made by means of restriction enzyme analysis,PCR and DNA sequencing.Results: The murine entire coding cDNA of 1592bp have been amplified from spleen cells.There was 99.8% homogenous nucleotide sequence as compared with standard sequence of T-bet from Genbank.Conclusion: We successfully cloned the cDNA of murine specific transcription factor T-bet.It will be the basis for studying immune interference of transferring gene.

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Available abstract

Objective: To clone all-length coding cDNA of specific transcription factor T-bet of murine T helper 1 type cells.Methods: Total RNA as well as mRNA were isolated from splenic cells of Balb/c mice and the entire coding cDNA sequence of T-bet was amplified by reverse transcription polymerase chain reaction(RT-PCR).The T-bet cDNA were cloned into the pGEM-T vector with a T base at 3' terminal.The identification was made by means of restriction enzyme analysis,PCR and DNA sequencing.Results: The murine entire coding cDNA of 1592bp have been amplified from spleen cells.There was 99.8% homogenous nucleotide sequence as compared with standard sequence of T-bet from Genbank.Conclusion: We successfully cloned the cDNA of murine specific transcription factor T-bet.It will be the basis for studying immune interference of transferring gene.

Key concepts: Complementary DNA, Molecular biology, Biology, Coding region, Rapid amplification of cDNA ends, Cloning (programming), Transcription factor, Gene

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