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2004•Journal of Branch Campus of the First Military MedicalRequires access

Cloning of human natural killer cell inhibitory receptor CD158b cDNA and its expression in E.coli

Cao Van Dong

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Abstract

Objective To clone and express the CD158b gene.Methods The gene which encodes matural killer cell inhibitory receptor was amplified by RT-PCR from peripheral blood mononuclear cells(PBMCs). Recombinant expression vector was constructed and sequenced after enzyme digestion. Results (1)Prospective amplified cDNA was obtained and the base sequence was identical with that recorded in GenBank after DNA sequencing. (2)Identification of pMBP-c-CD158b expression vector showed prospective result after the enzyme digestion analysis. (3)The size of recombinant protein was also coincident with the anticipotive aim and its proportion was 25% of the total protein. Conclusion CD158b cDNA was obtained by RT-PCR and successfully expressed in E.coli . The recombinant protein could provide an essential preparation for studying its functions.

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What this paper is about

Objective To clone and express the CD158b gene.Methods The gene which encodes matural killer cell inhibitory receptor was amplified by RT-PCR from peripheral blood mononuclear cells(PBMCs). Recombinant expression vector was constructed and sequenced after enzyme digestion. Results (1)Prospective amplified cDNA was obtained and the base sequence was identical with that recorded in GenBank after DNA sequencing. (2)Identification of pMBP-c-CD158b expression vector showed prospective result after the enzyme digestion analysis. (3)The size of recombinant protein was also coincident with the anticipotive aim and its proportion was 25% of the total protein. Conclusion CD158b cDNA was obtained by RT-PCR and successfully expressed in E.coli . The recombinant protein could provide an essential preparation for studying its functions.

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Available abstract

Objective To clone and express the CD158b gene.Methods The gene which encodes matural killer cell inhibitory receptor was amplified by RT-PCR from peripheral blood mononuclear cells(PBMCs). Recombinant expression vector was constructed and sequenced after enzyme digestion. Results (1)Prospective amplified cDNA was obtained and the base sequence was identical with that recorded in GenBank after DNA sequencing. (2)Identification of pMBP-c-CD158b expression vector showed prospective result after the enzyme digestion analysis. (3)The size of recombinant protein was also coincident with the anticipotive aim and its proportion was 25% of the total protein. Conclusion CD158b cDNA was obtained by RT-PCR and successfully expressed in E.coli . The recombinant protein could provide an essential preparation for studying its functions.

Key concepts: Complementary DNA, Recombinant DNA, Molecular biology, Cloning (programming), Biology, Peripheral blood mononuclear cell, GenBank, Gene

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