comConstruction and identification of a Mycobacterium smegmatis strain to targetedly deliver eukaryotic coexpression plasmid encoding human granulysin and murine single chain ILª²12

    YI Zhengª²Jun, ZHU Daoª²Yin, YANG Chun, LI Junª²Ming, HE Yongª²Lin, CHEN Quan

    Department of Microbiology and Immunology, Chongqing University of Medical Science, Chongqing 400016, China

    ¡¾Abstract¡¿ AIM: To construct a recombinant Mycobacterium smegmatis strain that can targetedly deliver the eukaryotic coexpression plasmid pZM03 encoding human granulysin (GLS) and murine single chain ILª²12 into macrophages. METHODS: Coding sequences of human granulysin, murine single chain ILª²12 and OriM were simultaneously cloned into pBudCE4.1, which had multiple promoters to form the shuttle plasmid pBudCE4.1/GLS/IL12/OriM (simply named pZM03). The shuttle plasmid was transformed into mc2ª²155 and identified from the recombinants by PCR. The targeting property of the recombinant was detected by coª²incubating the strains with murine macrophage RAW264.7. RESULTS: The new recombinant strain that could targetedly deliver pZM03 into macrophages was successfully constructed. CONCLUSION: It is feasible to targetedly deliver eukaryotic expression plasmids into macrophages by using Mycobacterium smegmatis as an attenuated biological vector. ......