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2005•Journal of Guangdong Medical CollegeRequires access

Cloning and eukaryotic expression of hpot1 cDNA in HeLa cells

Qifeng Zhu

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Abstract

Objective: To clone the whole length of coding sequence of hpot1(human protection of telomeres 1) cDNA,and construct an eukaryotic expression vector containing hpot1 to study the expression of hpot1 in HeLa cells.Methods:hpot1 cDNA was amplified with RT-PCR, directionally cloned into pcDNA3 plasmid and then confirmed by DNA sequencing;the recombinant pcDNA3-hpot1 plasmid was instantly transfected into HeLa cells;the expression of hpot1 was examined by RT-PCR and EMSA.Results:The recombinant pcDNA3-hpot1 plasmid was successfully constructed and the whole coding sequence and ORF of hpot1 extracted from the transfected Hela cells were proven to be correct.The transfected recombinant hpot1 plasmids was highly expressed in Hela cells after the transfection of 48 hours.Conclusion: The recombinant pcDNA3-hpot1 plasmid was successfully constructed and could be expressed in Hela cells.

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What this paper is about

Objective: To clone the whole length of coding sequence of hpot1(human protection of telomeres 1) cDNA,and construct an eukaryotic expression vector containing hpot1 to study the expression of hpot1 in HeLa cells.Methods:hpot1 cDNA was amplified with RT-PCR, directionally cloned into pcDNA3 plasmid and then confirmed by DNA sequencing;the recombinant pcDNA3-hpot1 plasmid was instantly transfected into HeLa cells;the expression of hpot1 was examined by RT-PCR and EMSA.Results:The recombinant pcDNA3-hpot1 plasmid was successfully constructed and the whole coding sequence and ORF of hpot1 extracted from the transfected Hela cells were proven to be correct.The transfected recombinant hpot1 plasmids was highly expressed in Hela cells after the transfection of 48 hours.Conclusion: The recombinant pcDNA3-hpot1 plasmid was successfully constructed and could be expressed in Hela cells.

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Available abstract

Objective: To clone the whole length of coding sequence of hpot1(human protection of telomeres 1) cDNA,and construct an eukaryotic expression vector containing hpot1 to study the expression of hpot1 in HeLa cells.Methods:hpot1 cDNA was amplified with RT-PCR, directionally cloned into pcDNA3 plasmid and then confirmed by DNA sequencing;the recombinant pcDNA3-hpot1 plasmid was instantly transfected into HeLa cells;the expression of hpot1 was examined by RT-PCR and EMSA.Results:The recombinant pcDNA3-hpot1 plasmid was successfully constructed and the whole coding sequence and ORF of hpot1 extracted from the transfected Hela cells were proven to be correct.The transfected recombinant hpot1 plasmids was highly expressed in Hela cells after the transfection of 48 hours.Conclusion: The recombinant pcDNA3-hpot1 plasmid was successfully constructed and could be expressed in Hela cells.

Key concepts: Plasmid, Recombinant DNA, HeLa, Complementary DNA, Molecular biology, Transfection, Biology, Cloning (programming)

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