Construction of Retrovirus Vector with Ubc9 Gene and RecombinantVirus Producing Cell Line

Objective: To establish a recombinant retrovirus vector containing Ubc9 gene and to establish a stable virus packaging cell line expressing Ubc9 effectively and stably.Methods: Ubc9 gene was amplified from the plasmid pcmv6-xl6-ubc9 by PCR technique and subcloned to the retroviral vector pMSCVneo to construct the recombined retrovirus vector.The recombinant plasmid was transfected into packaging cell line PT67 with PolyJetTM and the efficient virus-producing cell line PT67-Ubc9 was screened out following G418 selection and collected virus infected NIH/3T3 cells.Results: The recombinant retroviral vector pMSCV-Ubc9 was identified by restrictive analysis and DNA sequencing.A stable virus producing cell line was selected and the retrovirus was effectively transfected into NIH3T3 cells.Conclusion: The recombinant retroviral vector pMSCV-Ubc9 was constructed successfully.A stable viral producing cell line PT67-Ubc9 was selected and established.