African Journal of
Microbiology Research

  • Abbreviation: Afr. J. Microbiol. Res.
  • Language: English
  • ISSN: 1996-0808
  • DOI: 10.5897/AJMR
  • Start Year: 2007
  • Published Articles: 5233

Full Length Research Paper

Construction the recombinant BCG targeting delivering IprI into macrophages: A new strategy of vaccine against tuberculosis

Yuwei Wang1,2*, Lei Xu3, Li Zhang3, Yongling He3, Chun Yang3 and Ailong Huang1  
1Key Laboratory of Molecular Biology on Infectious Diseases, Ministry of Education, Chongqing Medical University, Chongqing, China. 2Department of Bio-medical Engineering, Chongqing Medical University, Chongqing, China. 3Department of Microbiology, Chongqing Medical University, Chongqing, China
Email: [email protected]

  •  Accepted: 27 August 2012
  •  Published: 12 February 2013


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