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目的:研究新发现的小鼠肿瘤/睾丸抗原Biot2对小鼠NIH3T3细胞生物学行为的影响,探讨基因的生物学功能。方法:构建真核表达质粒pcDNA3.1(+)-Biot2,由脂质体包裹稳定转染NIH3T3细胞,用Realtime RT-PCR、Western blot等方法筛选和鉴定Biot2表达阳性细胞克隆,研究其与细胞增殖相关的生物学特征的变化。结果:成功构建真核表达重组质粒pcDNA3.1(+)-Biot2,并稳定转染于NIH3T3细胞。与对照组相比,转染了Biot2cDNA的NIH3T3细胞生长速率明显增快。结论:成功地建立稳定转染小鼠新基因Biot2的NIH3T3细胞克隆;Biot2基因能影响细胞增殖的调节,促进NIH3T3细胞的增殖,为进一步研究基因生物学功能奠定了基础。
OBJECTIVE: To study the effect of newly discovered mouse tumor / testis antigen Biot2 on the biological behavior of mouse NIH3T3 cells and to explore the biological function of the gene. Methods: The eukaryotic expression plasmid pcDNA3.1 (+) - Biot2 was constructed and stably transfected into NIH3T3 cells by liposome. The positive clones of Biot2 positive cells were screened and identified by Realtime RT-PCR and Western blot. Changes in biological characteristics associated with proliferation. Results: The eukaryotic expression plasmid pcDNA3.1 (+) - Biot2 was successfully constructed and stably transfected into NIH3T3 cells. Compared with the control group, NIH3T3 cells transfected with Biot2 cDNA grew significantly faster. CONCLUSION: NIH3T3 cell clone stably transfected with Biot2, a novel gene of mouse, has been successfully established. Biot2 gene can affect the regulation of cell proliferation and promote the proliferation of NIH3T3 cells, which lays the foundation for further study of gene biology.