A lacZ Reporter-Based Strategy for Rapid Expression Analysis and Target Validation of Mycobacterium tuberculosis Latent Infection Genes

详细信息    查看全文

• 作者：Shivani Sood ; Satinder Kaur ; Rahul Shrivastava
• 刊名：Current Microbiology
• 出版年：2016
• 出版时间：February 2016
• 年：2016
• 卷：72
• 期：2
• 页码：213-219
• 全文大小：577 KB
• 参考文献：1.Betts JC, Lukey PT, Robb LC, McAdam RA, Duncan K (2002) Evaluation of a nutrient starvation model of Mycobacterium tuberculosis persistence by gene and protein expression profiling. Mol Microbiol 43:717–731PubMed CrossRef
  2.Carroll P, Schreuder LJ, Muwanguzi-Karugaba J, Wiles S, Robertson BD, Ripoll J, Ward TH, Bancroft GJ, Schaible UE, Parish T (2010) Sensitive detection of gene expression in mycobacteria under replicating and non-replicating conditions using optimized far-red reporters. PLoS ONE 5(3):e9823PubMed PubMedCentral CrossRef
  3.Cobat A, Orlova M, Barrera L, Schurr E (2013) Host genomics and control of tuberculosis infection. Public Health Genomics 16(1–2):44–49PubMed CrossRef
  4.Coralli C, Cemazar M, Kanthou C, Tozer GM, Dachs GU (2001) Limitations of the reporter green fluorescent protein under simulated tumor conditions. Cancer Res 61(12):4784–4790PubMed
  5.Davis SL, Be NA, Lamichhane G, Nimmagadda S, Pomper MG, Bishai WR, Jain SK (2009) Bacterial thymidine kinase as a non-invasive imaging reporter for Mycobacterium tuberculosis in live animals. PLoS ONE 4(7):e6297PubMed PubMedCentral CrossRef
  6.Deng Z, Shan Y, Pan Q, Gao X, Yan A (2013) Anaerobic expression of the gadE-mdtEF multidrug efflux operon is primarily regulated by the two-component system ArcBA through antagonizing the H-NS mediated repression. Front Microbiol. doi:10.​3389/​fmicb.​2013.​00194
  7.Dhandayuthapani S, Via L, Thomas C, Horowitz P, Deretic D, Deretic V (1995) Green fluorescent protein as a marker for gene expression and cell biology of mycobacterial interactions with macrophages. Mol Microbiol 17(5):901–912PubMed CrossRef
  8.Dick T, Lee BH, Murugasu-Oei B (1998) Oxygen depletion induced dormancy in Mycobacterium smegmatis. FEMS Microbiol Lett 163(2):159–164PubMed CrossRef
  9.Faye T, Asebo A, Salehian Z, Langsrud T, Nes IF, Brede DA (2008) Construction of a reporter vector system for in vivo analysis of promoter activity in Propionibacterium freudenreichii. Appl Environ Microbiol 74(11):3615–3617PubMed PubMedCentral CrossRef
  10.Hutter B, Dick T (1998) Increased alanine dehydrogenase activity during dormancy in Mycobacterium smegmatis. FEMS Microbiol Lett 167(1):7–11PubMed CrossRef
  11.Josserand V, Texier-Nogues I, Huber P, Favrot MC, Coll J-L (2007) Non-invasive in vivo optical imaging of the lacZ and luc gene expression in mice. Gene Ther 14(22):1587–1593PubMed CrossRef
  12.Mayuri Bagchi G, Das TK, Tyagi JS (2002) Molecular analysis of the dormancy response in Mycobacterium smegmatis: expression analysis of genes encoding the DevR-DevS two-component system, Rv3134c and chaperone alpha-crystallin homologues. FEMS Microbiol Lett 211(2):231–237PubMed
  13.McKinney JD, zu Bentrup KH, Muñoz-Elías EJ, Miczak A, Chen B, Chan W-T, Swenson D, Sacchettini JC, Jacobs WR, Russell DG (2000) Persistence of Mycobacterium tuberculosis in macrophages and mice requires the glyoxylate shunt enzyme isocitrate lyase. Nature 406(6797):735–738PubMed CrossRef
  14.Miller JH (1972) Experiments in molecular genetics. Cold Spring Harbor Laboratory, Cold Spring Harbor
  15.Murphy DJ, Brown JR (2007) Identification of gene targets against dormant phase Mycobacterium tuberculosis infections. BMC Infect Dis 7:84PubMed PubMedCentral CrossRef
  16.New D, Miller-Martini D, Wong Y (2003) Reporter gene assays and their applications to bioassays of natural products. Phytother Res 17(5):439–448PubMed CrossRef
  17.Pelicic V, Jackson M, Reyrat J-M, Jacobs WR, Gicquel B, Guilhot C (1997) Efficient allelic exchange and transposon mutagenesis in Mycobacterium tuberculosis. Proc of the Natl Acad of Sci U.S.A. 94(20):10955–10960CrossRef
  18.Saviola B (2013) Mycobacterium tuberculosis adaptation to survival in a human host. In: Mahboub BH, Vats MG (eds.) Tuberculosis - current issues in diagnosis and management. INTECH Open Acess Publisher, Croatia, European Union, p 3–18
  19.Shi L, Jung Y-J, Tyagi S, Gennaro ML, North RJ (2003) Expression of Th1-mediated immunity in mouse lungs induces a Mycobacterium tuberculosis transcription pattern characteristic of nonreplicating persistence. Proc Natl Acad Sci USA 100(1):241–246PubMed PubMedCentral CrossRef
  20.Stewart GR, Newton SM, Wilkinson KA, Humphreys IR, Murphy HN, Robertson BD, Wilkinson RJ, Young DB (2005) The stress-responsive chaperone α-crystallin 2 is required for pathogenesis of Mycobacterium tuberculosis. Mol Microbiol 55(4):1127–1137PubMed CrossRef
  21.Stover CK, de la Cruz VF, Fuerst TR, Burlein JE, Benson LA, Bennett LT, Bansal GP, Young JF, Lee MH, Hatfull GF, Snapper SB, Barletta RG, Jacobs WR, Bloom BR (1991) New use of BCG for recombinant vaccines. Nature 351(6326):456–460PubMed CrossRef
  22.Tsien RY (1998) The green fluorescent protein. Annu Rev Biochem 67(1):509–544PubMed CrossRef
  23.Villemagne B, Crauste C, Flipo M, Baulard AR, Déprez B, Willand N (2012) Tuberculosis: the drug development pipeline at a glance. Eur J Med Chem 51:1–16PubMed CrossRef
  24.Voskuil MI, Visconti K, Schoolnik G (2004) Mycobacterium tuberculosis gene expression during adaptation to stationary phase and low-oxygen dormancy. Tuberculosis 84(3):218–227PubMed CrossRef
  25.Wayne LG, Hayes LG (1996) An in vitro model for sequential study of shiftdown of Mycobacterium tuberculosis through two stages of nonreplicating persistence. Infect Immun 64(6):2062–2069PubMed PubMedCentral
  26.WHO. Global tuberculosis report (2014) World Health Organization Geneva; http://​www.​who.​int/​tb/​publications/​global_​report/​en/​ . Accessed 8 Aug 2015
  27.Zahrt TC (2003) Molecular mechanisms regulating persistent Mycobacterium tuberculosis infection. Microbes Infect 5(2):159–167PubMed CrossRef
  28.Zhang Y (2004) Persistent and dormant tubercle bacilli and latent tuberculosis. Front Biosci 9:1136–1156PubMed CrossRef
  29.Zhang Y, Xiao M, Horiyama T, Zhang Y, Li X, Nishino K, Yan A (2011) The multidrug efflux pump MdtEF protects against nitrosative damage during the anaerobic respiration in Escherichia coli. J Biol Chem 286(30):26576–26584PubMed PubMedCentral CrossRef
  30.Zumla A, Atun R, Maeurer M, Kim PS, Jean-Philippe P, Hafner R, Schito M (2012) Eliminating tuberculosis and tuberculosis–HIV co-disease in the 21st century: key perspectives, controversies, unresolved issues, and needs. J Infect Dis 205(suppl 2):S141–S146PubMed PubMedCentral CrossRef
  
• 作者单位：Shivani Sood (1)
  Satinder Kaur (2)
  Rahul Shrivastava (1)
  
  1. Department of Biotechnology and Bioinformatics, Jaypee University of Information Technology, Waknaghat, Solan, Himachal Pradesh, 173234, India
  2. Division of Microbiology, Central Drug Research Institute, Lucknow, Uttar Pradesh, 226001, India
  
• 刊物类别：Biomedical and Life Sciences
• 刊物主题：Life Sciences
  Microbiology
  Biotechnology
  
• 出版者：Springer New York
• ISSN：1432-0991

文摘

We report a novel lacZ fusion vector and demonstrate its utility for expression analysis of genes associated with Mycobacterium tuberculosis latent infection. The vector contains E. coli (oriE) and mycobacterial (oriM) origins of replication, a kanamycin resistance gene (Kmr) as selection marker, and a lacZ reporter gene in fusion with MCS for cloning of upstream regulatory sequence of the desired genes. β-galactosidase activity of the vector was standardized for expression analysis under latent mycobacterial conditions using Phsp60, a constitutive mycobacterial promoter, utilizing Mycobacterium smegmatis as model organism. Validation of the vector was done by cloning and expression analysis of PhspX (alpha crystalline) and Picl (isocitrate lyase), promoters from two of the genes shown to be involved in M. tuberculosis persistence. Both genes showed appreciable levels of β-galactosidase expression under hypoxia-induced persistent conditions in comparison to their actively replicating state. Expression analysis of a set of hypothetical genes was also done, of which Rv0628c showed increased expression under persistent conditions. The reported fusion vector and the strategy can be effectively used for short listing and validation of drug targets deduced from various non-conclusive approaches such as bioinformatics and microarray analysis against latent/persistent form of mycobacterial infection.