Construction and expression of recombinant bacille calmette-guerin carrying murine interleukin-12 / 重庆医科大学学报

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.