ZHANG Ruhua, XU Shuangbing, WU Yuanzhong, et al. Construction and Identification of hSSB1 Retrovirus Expressing Vector and Screening of Stable Transfected Cells[J]. Acta Scientiarum Naturalium Universitatis SunYatseni, 2012,51(2):73-76.
ZHANG Ruhua, XU Shuangbing, WU Yuanzhong, et al. Construction and Identification of hSSB1 Retrovirus Expressing Vector and Screening of Stable Transfected Cells[J]. Acta Scientiarum Naturalium Universitatis SunYatseni, 2012,51(2):73-76.DOI:
hSSB1 is a key signaling moleculue that participates in DNA damage response. In this study
the cDNA of
hSSB1
gene amplified by RT-PCR was inserted into the retroviral vector pBABE. The recombinant positive plasmid clone was identified by endonuclease digestion
PCR amplification and sequencing analysis. The retroviral expression vector pBABE-
hSSB1
and PIK packaging plasmid were cotransfected into 293T cells to produce the retrovirus. The packaging retrovirus was then infected into cancer cells and the overexpression cells were selected with puromycin. pBABE-
hSSB1
positive clones have been validated to be correct by restriction endonuclease
PCR amplification and DNA sequencing analysis. The protein level of hSSB1 in pBABE-
hSSB1
transfected cancer cell line was significantly up-regulated as validated by Western blotting. Our data indicate that the recombinant plasmid of pBABE-
hSSB1
was successfully constructed and transfected stably into cancer cells. It established a favorable foundation for further functional study.