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Cloning,sequencing and expression of human bit1 cDNA gene

Zhe Wang

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Abstract

AIM: To clone the human bit1 cDNA gene and express it in E. coli. METHODS: Total RNA was extracted from fresh placenta tissue and reverse transcripted to single chain cDNA. By using it as the template, the human bit1 cDNA gene was amplified. The cDNA gene was then cloned into pUC19. After the sequence was verified, the cDNA gene was subcloned into pGEX-4T3 for expression. RESULTS: The human bit1 cDNA gene was amplified by using the designed primer and the gene was expressed in E.coli induced by IPTG. The expressed protein accounted for 40.6% of the total amount of protein. CONCLUSION: The human bit1 cDNA gene is amplified and expressed in E.coli, which is important for the study on the function of human bit1 protein.

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What this paper is about

AIM: To clone the human bit1 cDNA gene and express it in E. coli. METHODS: Total RNA was extracted from fresh placenta tissue and reverse transcripted to single chain cDNA. By using it as the template, the human bit1 cDNA gene was amplified. The cDNA gene was then cloned into pUC19. After the sequence was verified, the cDNA gene was subcloned into pGEX-4T3 for expression. RESULTS: The human bit1 cDNA gene was amplified by using the designed primer and the gene was expressed in E.coli induced by IPTG. The expressed protein accounted for 40.6% of the total amount of protein. CONCLUSION: The human bit1 cDNA gene is amplified and expressed in E.coli, which is important for the study on the function of human bit1 protein.

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Available abstract

AIM: To clone the human bit1 cDNA gene and express it in E. coli. METHODS: Total RNA was extracted from fresh placenta tissue and reverse transcripted to single chain cDNA. By using it as the template, the human bit1 cDNA gene was amplified. The cDNA gene was then cloned into pUC19. After the sequence was verified, the cDNA gene was subcloned into pGEX-4T3 for expression. RESULTS: The human bit1 cDNA gene was amplified by using the designed primer and the gene was expressed in E.coli induced by IPTG. The expressed protein accounted for 40.6% of the total amount of protein. CONCLUSION: The human bit1 cDNA gene is amplified and expressed in E.coli, which is important for the study on the function of human bit1 protein.

Key concepts: Complementary DNA, Biology, Gene, Molecular biology, Rapid amplification of cDNA ends, Gene expression, Expressed sequence tag, pUC19

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