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《Acta Academiae Medicine Militaris Tertiae》 2002-01
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Transfection of recombined rat IFN-γ into rat alveolar macrophage and airway epithelial cells in vitro

HUANG Gui jun, LONG Jiang, MA Zhuang, QIAN Gui sheng, LI Shu ping, CHEN Wei zhong (Institute of Respiratory Diseases, Xingqiao Hospital, Third Military Medical University, Chongqing 400037,China)  
Objective To recombine IFN γ expression vector and study its expression in alveolar macrophage (AM) and airway epithelial cells of Wistar rats after transfection in vitro . Methods IFN γ gene was cloned from Wistar rats with RT PCR, and then inserted into pLXSN vector with DNA sequencing. The vector was transfected into AM and airway epithelial cells in cell culture by using cationic liposomes. The integration of pLXSN IFN and pLXSN with the genomic DNA of the transfected cells was detected with PCR. And the expression level and activity of IFN γ in the supernatant were investigated with ELISA and L929 cytopathogenicity inhibition assay. Results DNA sequencing proved that the sequence of the cloned IFN γgene was identical to its counterpart in GenBank and the recombinant pLXSN IFN expression vector was successfully constructed. A PCR product of 620 bp in the cells with pLXSN IFN transfection and one of 140 bp in pLXSN transfected cells were demonstrated in agrose gel with eletrophoresis. The levels and activity of IFN γ in the culture supernatant after pLXSN IFN transfection were markedly higher than those of the cells transfected with pLXSN ( P 0.01). Conclusion The recombinant IFN γ gene expression vector is successfully transfected into AM and airway epithelial cells in vitro . It can integrate with the genomic DNA of the cells, and express active IFN γ, which is useful for further investigation in vivo .
【Fund】: 全军“十五”攻关课题面上项目 (0 1MB0 0 1)
【CateGory Index】: R346
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【Co-citations】
Chinese Journal Full-text Database 4 Hits
1 MA Zhuang, QIAN Gui sheng, HUANG Gui jun, MAO Bao lin (Institute of Respiratory Diseases, Xinqiao Hospital, Third Military Medical University, Chongqing 400037, China);Effect of IFN-γ inhalation on some cytokines of immunocompromised rats[J];Acta Academiae Medicine Militaris Tertiae;2001-02
2 Zhuo Anshan Chen Aijuan Shen Fanxing Cheng Guozun Huang Qiang (465th hospital of P L A JiLin 132011);Effect of Various Immunomodulators on the Prodution of Anti SRBC Antibody and IFN γ by Splenocytes in Rats with Burn[J];BASIC MEDICAL SCIENCES AND CLINICS;1998-03
3 Ma Zhuang, Qian Guisheng, Huang Guijun. Xinqiao Hospital, Third Military Medical University, Chongqing 400037;STUDY OF INCREASING THE LEVEL OF IFN-γ IN THE LUNGS OF IMMUNOCOMPROMISED RATS WITH GENE TRANSFECTION[J];Medical Journal of Chinese People's Liberation Army;2002-02
4 MA Zhuang, QIAN Gui sheng, HUANG Gui jun (Institute of Respiratory Medicine, Xinqiao Hospital, Third Mil itary Medical University, Chongqing 400037, China);Study of IFN-γ gene of rats transfected into airway epithelial cell of rats[J];Immunological Journal;2001-05
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