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ISSN 1001-5256 (Print)
ISSN 2097-3497 (Online)
CN 22-1108/R
Issue 6
Jun.  2014
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Construction of lentiviral vector encoding CLEC4M and overexpression of CLEC4M in K- 562 cells

DOI: 10.3969/j.issn.1001-5256.2014.06.010
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  • Received Date: 2013-09-18
  • Published Date: 2014-06-20
  • Objective To construct the lentiviral vector encoding CLEC4M and prepare K- 562 cells with stable overexpression of CLEC4M. Methods The gene sequence of normal CLEC4M was cloned by reverse transcription PCR and then inserted into GV166 vector to construct GV166- CLEC4M lentiviral expression vector, and then lentiviral packaging was performed by transfection of 293T cells. The obtained lentiviral liquid was used to infect human leukemia cell line K- 562. Real- time PCR and Western blot were used to detect the overexpression of CLEC4M in K- 562 cells. Results Sequencing showed that the recombinant lentiviral expression plasmid GV166- CLEC4M was successfully constructed. Lentiviruses could efficiently infect K- 562 cells, according to real- time PCR. CLEC4M was successfully over- expressed in K- 562 cells at mRNA and protein levels. Conclusion The construction of lentiviral vector encoding CLEC4M lays a foundation for further study of CLEC4M gene involved in HCV entry into host cells.

     

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