【摘要】 目的:构建携带小鼠白细胞介素-12基因的重组卡介苗(Bacille calmette-guerin,BCG),并鉴定该重组菌向真核细胞递呈质粒的能力。方法:以分子生物学方法构建携带小鼠IL-12基因和分支杆菌复制子OriM的真核穿梭共表达质粒pBM12,以电转化的方式将pBM12质粒转入BCG构建重组BCG,抽提质粒进行PCR鉴定重组菌;将重组菌感染巨噬细胞,通过RT-PCR了解重组菌向真核细胞递呈质粒的能力。结果:电转化方式可将pBM12导入BCG,抽提质粒鉴定出与mIL-12基因相符的目的条带。以该重组BCG感染小鼠巨噬细胞,96h后用RT-PCR法也扩增出1632bp左右的基因条带。结论:成功构建含pBM12的重组BCG。该重组BCG能向小鼠巨噬细胞递呈携带基因。更多还原

【Abstract】 Objective:To construct recombinant bacille calmette-guerin (BCG) carrying murine interleukin-12 (mIL-12) genes,and to identify the ability of recombinant BCG for submitting plasmids to eukaryotic cells. Methods:Co-expression shuttle plasmid of pBM12, including mIL-12 gene and mycobacterial replicon of Orim,was built using molecular biological approach.The pBM12 was transfected to BCG competence by electroporation to construct recombinant BCG. The plasmid of recombinant BCG was extracted for PCR identification. The ability of submitting plasmids to eukaryotic cells was determined by RT-PCR. Results:The plasmid pBM12 may be transduced into BCG with electroporation. The specific bands for mIL-12 of the recombinant BCG were detected by PCR. The specific bands for 1632 bp mIL-12 were amplified from transfected macrophages after 96 hours byRT-PCR. Conclusion:The recombinant BCG carrying pBM12 was constructed successfully. The recombinant BCG can submit the carried genes to mice macrophages.更多还原