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ENGLISH ABSTRACT
JOURNAL ARTICLE
[Expression of FasL gene in porcine chondrocytes].
Xi Bao Yu Fen Zi Mian Yi Xue za Zhi = Chinese Journal of Cellular and Molecular Immunology 2005 March
AIM: To express the Fas ligand(FasL) gene in porcine chondrocytes.
METHODS: The porcine FasL gene fragment was amplified by RT-PCR and then inserted into the pGCEN retroviral vector. The recombinant vector was transfected into packaging cells PA317 which were then screened with G418. Supernatant of screened PA317 cells containing high titer recombinant virus was used to infect porcine chondrocytes. Expression of FasL in chondrocytes was analyzed by FACS and Western blot.
RESULTS: The recombinant pGCEN-FasL retroviral expression vector was successfully constructed as shown by restriction enzyme digestion analysis and DNA sequencing. FasL was expressed in 57% of infected chondrocytes. Western blot analysis also confirmed the expression of FasL in chondrocytes. The expressed FasL could induce apoptosis of Fas+ cells.
CONCLUSION: The recombinant pGCEN-FasL retroviral vector has been successfully constructed and FasL with biological activity was highly expressed in porcine chondrocytes, which lays the foundation for allogenic chondrocytes transplantation.
METHODS: The porcine FasL gene fragment was amplified by RT-PCR and then inserted into the pGCEN retroviral vector. The recombinant vector was transfected into packaging cells PA317 which were then screened with G418. Supernatant of screened PA317 cells containing high titer recombinant virus was used to infect porcine chondrocytes. Expression of FasL in chondrocytes was analyzed by FACS and Western blot.
RESULTS: The recombinant pGCEN-FasL retroviral expression vector was successfully constructed as shown by restriction enzyme digestion analysis and DNA sequencing. FasL was expressed in 57% of infected chondrocytes. Western blot analysis also confirmed the expression of FasL in chondrocytes. The expressed FasL could induce apoptosis of Fas+ cells.
CONCLUSION: The recombinant pGCEN-FasL retroviral vector has been successfully constructed and FasL with biological activity was highly expressed in porcine chondrocytes, which lays the foundation for allogenic chondrocytes transplantation.
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