小鼠白细胞介素12基因转染及其在黑色素瘤细胞B16F10中的表达

Transfection and expression of murine interleukin-12 gene in B16F10 melanoma cells

目的 :探索小鼠白细胞介素 12 (mIL - 12 )基因在小鼠黑色素瘤B16F10细胞中的表达。方法 :应用DNA重组技术将mIL - 12基因插入pcDNA3.1真核表达载体中 ,通过电穿孔转染B16F10细胞 ,筛选出阳性细胞克隆后 ,应用PCR、RT -PCR及Westernblot技术检测mIL - 12基因在B16F10细胞中的整合及表达。结果 :在DNA、mRNA及蛋白质 3个水平均证实mIL - 12基因已转染到B16F10细胞中并表达。结论 :mIL - 12基因可成功地转染体外培养的B16F10细胞并表达 ,为进一步研究IL - 12基因修饰的肿瘤细胞的基因瘤苗奠定了基础。

AIM: To study the expression of murine interleukin-12(mIL-12) gene in B16F10 melanoma cells. METHODS: mIL-12 gene was inserted into pcDNA3.1 eukaryotic expression vector by DNA recombination and then transfected into B16F10 melanoma cells by electroporation. The integration and expression of mIL-12 gene in B16F10 melanoma cells were detected by PCR,RT-PCR and Western blot after positive cell clones had been screened. RESULTS: mIL-12 gene was confirmed to have been transfected and expressed in B16F10 cells on three levels of DNA, mRNA and protein. CONCLUSION: mIL-12 gene can successfully be transfected and expressed in B16F10 melanoma cells in vitro , which lays a foundation for the further investigation of mIL-12 gene tumor vaccine.