Abstract

This study was aimed at constructing a recombinant Bacillus Calmette-Guérin (BCG) thatcan target the delivery of the intracellular pathogen resistance  = 1 \* ROMAN I (Ipr1) gene into macrophages. To achieve this, a eukaryotic plasmid pBGOI was constructed, which co-expressed Ipr1 and green fluorescent protein (GFP). Then, pBGOI was transfected into murine macrophage cell line RAW264.7 and transformed into BCG to construct the recombinant BCG, which was used to immunize C3HeB/FeJ mice intranasally. Ipr1expression in vitro and vivo was detected by real time-polymerase chain reaction (RT-PCR), fluorescence microscope, Western-blot and immunohistochemistry. Results on restriction enzyme digestion and sequence analysis showed that pBGOI had been constructed successfully. Ipr1 expression was detected not only in macrophages infected with pBGOI, but also in lung and spleen tissues of C3HeB/FeJ mice that were immunized by recombinant BCG. This study therefore provides a good basis for further research on the function and mechanisms of Ipr1 against tuberculosis.

Key words: Ipr1 gene, Bacillus Calmette-Guérin (BCG), macrophage, Mycobacterium tuberculosis, vaccin