Construction and expression of retrovirus vector harboring genes of HIVª²1 coreceptors ligandsª²RANTES and SDFª²1

    ZHANG Ying, WANG Jiuª²Ping, LI Jinª²Ge, WANG Pingª²Zhong, CHEN Weiª²Hong, BAI Xueª²Fan

    Center of Diagnosis & Treatment for Infectious Diseases of PLA, Tangdu Hospital, Fourth Military Medical University, Xi¬ðan 710038, China

    ¡¾Abstract¡¿ AIM£º To construct the retrovirus vector of HIVª²1 coreceptors ligandsª²RANTES and SDFª²1 and to identify their expression. METHODS: The genes of RANTES and SDFª²1 were reconstructed in retrovirus vector and transferred into packaging cell Bing by DNAª²calcium phosphate coprecipitation. The clones of the cells transferred with RANTES and SDFª²1 were selected by G418. Targeted NIH3T3 cells were infected with the virus granules secreting from Bing and also selected by G418. Indirect immumofluorescence was used to confirm the expression of RANTES and SDFª²1 in NIH3T3 cells. RESULTS: The reconstruction of pLNCXª²Rª²Kª²Sª²K was successful. Positive clones which strongly resisted G418 were selected and retrovirus granules with high biological activity were prepared. RANTES and SDFª²1 were observed to have expressed in NIH3T3 cells by indirect immumofluorescence. CONCLUSION: Retrovirus vector pLNCXª²Rª²Kª²Sª²K is constructed and infects NIH3T3 cells successfully, which will help further research of gene therapy for HIVª²1 infection. ......