Abstract:
:This study describes the use and the advantages of the green fluorescent protein (GFP) as a reporter molecule for mycobacteria. The gfp gene from Aequorea victoria was placed under the control of the hsp60 promoter in the shuttle vector pGFM-11. The gfp expression in the recombinant Mycobacterium smegmatis and BCG was readily detected on agar plates by the development of an intense green fluorescence upon irradiation with long-wave u.v. light. In mycobacteria containing a pGFM-11 derivative that lacks the hsp60 promoter, no fluorescence was observed. However, this plasmid was successfully used as a promoter-probe vector to identify BCG promoters. The fluorescence emission of GFP in mycobacteria harbouring pGFM-11 and grown in liquid media could be quantified by spectrofluorimetry. This allowed for easy assessment of drug susceptibility. As GFP does not require the addition of substrates or co-factors, the green fluorescent bacilli could be directly observed within infected macrophages using fluorescence and laser confocal microscopy, or in tissue sections of infected mice. Finally, infected cells or free-living recombinant mycobacteria could also be analysed by flow cytometry. The GFP thus appears to be a convenient reporter for mycobacteria, allowing tracing of recombinant mycobacteria, isolation of promoters with interesting properties, in vivo drug testing and the development of new diagnostic tools.
journal_name
Mol Microbioljournal_title
Molecular microbiologyauthors
Kremer L,Baulard A,Estaquier J,Poulain-Godefroy O,Locht Cdoi
10.1111/j.1365-2958.1995.mmi_17050913.xsubject
Has Abstractpub_date
1995-09-01 00:00:00pages
913-22issue
5eissn
0950-382Xissn
1365-2958journal_volume
17pub_type
杂志文章abstract::Bacterial cytokinesis requires the divisome, a complex of proteins that co-ordinates the invagination of the cytoplasmic membrane, inward growth of the peptidoglycan layer and the outer membrane. Assembly of the cell division proteins is tightly regulated and the order of appearance at the future division site is well...
journal_title:Molecular microbiology
pub_type: 杂志文章
doi:10.1111/j.1365-2958.2008.06141.x
更新日期:2008-04-01 00:00:00
abstract::Virion proteins recognize their cognate nucleic acid for encapsidation into virions through recognition of a specific nucleotide sequence contained within that nucleic acid. Viruses like bacteriophage P22, which have partially circularly permuted, double-stranded virion DNAs, encapsidate DNA through processive series ...
journal_title:Molecular microbiology
pub_type: 杂志文章
doi:10.1046/j.1365-2958.2002.03114.x
更新日期:2002-09-01 00:00:00
abstract::N-glycosylation in the endoplasmic reticulum is an essential protein modification and highly conserved in evolution from yeast to man. Defects of N-glycosylation in humans lead to congenital disorders. The pivotal step of this pathway is the transfer of the evolutionarily conserved lipid-linked core-oligosaccharide to...
journal_title:Molecular microbiology
pub_type: 杂志文章
doi:10.1111/j.1365-2958.2005.04981.x
更新日期:2006-02-01 00:00:00
abstract::We have previously described HpuAB, a two-component receptor that mediates binding to haemoglobin (Hb), haemoglobin-haptoglobin (Hb-Hp) and apo-haptoglobin (Hp). In this communication, we constructed non-polar mutations in the hpuA and hpuB loci to examine the individual roles of HpuA and HpuB. Our results indicate th...
journal_title:Molecular microbiology
pub_type: 杂志文章
doi:10.1046/j.1365-2958.1999.01409.x
更新日期:1999-06-01 00:00:00
abstract::The four motor proteins PomA, PomB, MotX and MotY, which are believed to be stator proteins, are essential for motility by the Na(+)-driven flagella of Vibrio alginolyticus. When we purified the flagellar basal bodies, MotX and MotY were detected in the basal body, which is the supramolecular complex comprised of the ...
journal_title:Molecular microbiology
pub_type: 杂志文章
doi:10.1111/j.1365-2958.2006.05435.x
更新日期:2006-11-01 00:00:00
abstract::The wrinkly spreader (WS) genotype of Pseudomonas fluorescens SBW25 colonizes the air-liquid interface of spatially structured microcosms resulting in formation of a thick biofilm. Its ability to colonize this niche is largely due to overproduction of a cellulosic polymer, the product of the wss operon. Chemical analy...
journal_title:Molecular microbiology
pub_type: 杂志文章
doi:10.1046/j.1365-2958.2003.03670.x
更新日期:2003-10-01 00:00:00
abstract::This past January, in Cuernavaca Mexico, a conglomerate of scientists met to discuss the contemporary view of Bacterial Locomotion and Signal Transduction (BLAST). The BLAST meetings represent a field that has its roots in chemotaxis and the flagellum-based motility but now encompass all types of cellular movement and...
journal_title:Molecular microbiology
pub_type: 杂志文章
doi:10.1111/j.1365-2958.2009.06742.x
更新日期:2009-07-01 00:00:00
abstract::The Helicobacter pylori vacuolating cytotoxin (VacA) induces degenerative vacuolation of sensitive mammalian cell lines. Although evidence is accumulating that VacA enters cells and functions from an intracellular site of action, the biochemical mechanism by which VacA mediates cellular vacuolation has not been establ...
journal_title:Molecular microbiology
pub_type: 杂志文章
doi:10.1046/j.1365-2958.2002.02818.x
更新日期:2002-03-01 00:00:00
abstract::Although diminutive in size, bacteria possess highly diverse and spatially confined cellular structures. Two related alphaproteobacteria, Sinorhizobium meliloti and Caulobacter crescentus, serve as models for investigating the genetic basis of morphological variations. S. meliloti, a symbiont of leguminous plants, syn...
journal_title:Molecular microbiology
pub_type: 杂志文章
doi:10.1111/j.1365-2958.2012.08064.x
更新日期:2012-06-01 00:00:00
abstract::Francisella tularensis are the causative agent of the zoonotic disease, tularaemia. Among four F. tularensis subspecies, ssp. novicida (F. novicida) is pathogenic only for immunocompromised individuals, while all four subspecies are pathogenic for mice. This study utilized proteomic and bioinformatic approaches to ide...
journal_title:Molecular microbiology
pub_type: 杂志文章
doi:10.1111/j.1365-2958.2006.05365.x
更新日期:2006-10-01 00:00:00
abstract::Of the four Plasmodium species that routinely cause malaria in humans, Plasmodium falciparum is responsible for the majority of malaria mortality and consequently gets most of the headlines. Outside Africa, however, more malaria cases are caused by its distant cousin Plasmodium vivax, resulting in a daunting morbidity...
journal_title:Molecular microbiology
pub_type: 杂志文章
doi:10.1111/j.1365-2958.2005.04518.x
更新日期:2005-03-01 00:00:00
abstract::The human stomach pathogen Helicobacter pyloriattaches to healthy and inflamed gastric tissue through members of a paralogous family of 'Helicobacter outer membrane proteins' (Hops), including adhesins BabA, SabA, HopQ, LabA and HopZ. Hops share a conserved 25 kDa C-terminal region that is thought to form an autotrans...
journal_title:Molecular microbiology
pub_type: 杂志文章
doi:10.1111/mmi.14075
更新日期:2018-10-01 00:00:00
abstract::Malaria-causing parasites are transmitted from humans to mosquitoes when developmentally arrested gametocytes are taken up by a female Anopheles during a blood meal. The changes in environment from human to mosquito activate gametogenesis, including a drop in temperature, a rise in pH, and a mosquito-derived molecule,...
journal_title:Molecular microbiology
pub_type: 杂志文章,评审
doi:10.1111/mmi.14633
更新日期:2020-10-28 00:00:00
abstract::Tuberculosis remains the greatest cause of death worldwide due to a single pathogen. In order to identify the genes required for the pathogenicity of Mycobacterium tuberculosis, a functional genomic approach was developed. A library of signature-tagged transposon mutants of this bacterium was constructed and screened ...
journal_title:Molecular microbiology
pub_type: 杂志文章
doi:10.1046/j.1365-2958.1999.01593.x
更新日期:1999-10-01 00:00:00
abstract::The Pseudomonas aeruginosa algD gene is the first gene of an operon encoding most of the enzymes necessary for biosynthesis of the exopolysaccharide alginate. Transcriptional activation of algD results in the high-level synthesis of alginate, an important P. aeruginosa virulence factor with antiphagocytic and adherenc...
journal_title:Molecular microbiology
pub_type: 杂志文章
doi:10.1046/j.1365-2958.1999.01550.x
更新日期:1999-09-01 00:00:00
abstract::Lipopolysaccharides (LPS), particularly the O-antigen component, are one of many virulence determinants necessary for Shigella flexneri pathogenesis. O-antigen biosynthesis is determined mostly by genes located in the rfb region of the chromosome. The rfc/wzy gene encodes the O-antigen polymerase, an integral membrane...
journal_title:Molecular microbiology
pub_type: 杂志文章
doi:10.1046/j.1365-2958.1998.00884.x
更新日期:1998-06-01 00:00:00
abstract::The ability of Salmonella enterica serovar Typhimurium to cause disease depends upon the co-ordinated expression of many genes located around the Salmonella chromosome. Specific pathogenicity loci, termed Salmonella pathogenicity islands, have been shown to be crucial for the invasion and survival of Salmonella within...
journal_title:Molecular microbiology
pub_type: 杂志文章
doi:10.1046/j.1365-2958.2001.02192.x
更新日期:2001-01-01 00:00:00
abstract::An msDNA operon, consisting of genes for msDNA and a reverse transcriptase, is present in Escherichia coliB and absent from E. coliK12. We have found that the msDNA operon is located on a DNA fragment, longer than 15kb, that is absent from E. coliK12. Using conjugation, P1 transduction, and nucleic acid hybridization ...
journal_title:Molecular microbiology
pub_type: 杂志文章
doi:10.1111/j.1365-2958.1990.tb00582.x
更新日期:1990-12-01 00:00:00
abstract::Chromalveolates like the diatom Phaeodactylum tricornutum arose through the uptake of a red alga by a phagotrophic protist, a process termed secondary endosymbiosis. In consequence, the plastids are surrounded by two additional membranes compared with primary plastids. This plastid morphology poses additional barriers...
journal_title:Molecular microbiology
pub_type: 杂志文章
doi:10.1111/j.1365-2958.2010.07142.x
更新日期:2010-05-01 00:00:00
abstract::The development of new drugs for treating Pneumocystis carinii infections in AIDS patients is hampered by the lack of long-term culture systems, and by our generally limited knowledge of this organism. Recently, however, we observed significant activity of various benzimidazoles against growth of this organism in shor...
journal_title:Molecular microbiology
pub_type: 杂志文章
doi:10.1111/j.1365-2958.1992.tb02204.x
更新日期:1992-11-01 00:00:00
abstract::F plasmid traY protein binding to wild-type or deleted regions containing the TraY-binding site, sbyA, was studied in vitro. The principal DNA-protein complex was formed with DNA segments including the sbyA site defined by footprinting and (with lesser affinity) with truncated segments that retained the leftward two-t...
journal_title:Molecular microbiology
pub_type: 杂志文章
doi:10.1111/j.1365-2958.1994.tb00327.x
更新日期:1994-02-01 00:00:00
abstract::Temperate phage P2 has the capacity to function as a helper for the defective, unrelated, satellite phage P4. In the absence of a helper, P4 can either lysogenize its host or establish itself as a plasmid. For lytic growth, P4 requires the structural genes, packaging and lysis functions of the helper. P4 can get acces...
journal_title:Molecular microbiology
pub_type: 杂志文章
doi:10.1046/j.1365-2958.1998.01132.x
更新日期:1998-12-01 00:00:00
abstract::The suf and isc operons of Escherichia coli have been implicated in Fe-S cluster assembly. However, it has been unclear why E. coli has two systems for Fe-S cluster biosynthesis. We have examined the regulatory characteristics and mutant phenotypes of both operons to discern if the two operons have redundant functions...
journal_title:Molecular microbiology
pub_type: 杂志文章
doi:10.1111/j.1365-2958.2004.04025.x
更新日期:2004-05-01 00:00:00
abstract::In the budding yeast Saccharomyces cerevisiae, the preservation of the mating competent haploid (a or alpha) and the mating incompetent diploid (a/alpha) is necessary to prevent aneuploidy. Once haploid cells respond to pheromone, the mating-specific signal transduction pathway is activated, and the MAP kinase Fus3 ph...
journal_title:Molecular microbiology
pub_type: 杂志文章
doi:10.1046/j.1365-2958.2002.03213.x
更新日期:2002-11-01 00:00:00
abstract::The Dot/Icm system is a type IVb secretion system used by Legionella pneumophila to modulate vesicular transport in both protozoan and mammalian host cells. It has been shown that proteins and processes that are highly conserved in all eukaryotic cells are targets for some of the proteins injected by the Dot/Icm syste...
journal_title:Molecular microbiology
pub_type: 杂志文章
doi:10.1111/j.1365-2958.2005.04595.x
更新日期:2005-05-01 00:00:00
abstract::A Lactobacillus casei BL23 strain defective in an OmpR-family response regulator encoded by LCABL_18980 (PrcR, RR11), showed enhanced proteolytic activity caused by overexpression of the gene encoding the proteinase PrtP. Transcriptomic analysis revealed that, in addition to prtP expression, PrcR regulates genes encod...
journal_title:Molecular microbiology
pub_type: 杂志文章
doi:10.1111/mmi.13299
更新日期:2016-04-01 00:00:00
abstract::The gene cbhA from the cellulolytic bacterium Cellulomonas fimi encodes a protein of 872 amino acids designated cellobiohydrolase A (CbhA). Mature CbhA contains 832 amino acid residues and has a predicted molecular mass of 85,349 Da. It is composed of five domains: an N-terminal catalytic domain, three repeated sequen...
journal_title:Molecular microbiology
pub_type: 杂志文章
doi:10.1111/j.1365-2958.1994.tb01030.x
更新日期:1994-05-01 00:00:00
abstract::The outer membrane channel TolC is a key component of multidrug efflux and type I secretion transporters in Escherichia coli. Mutational inactivation of TolC renders cells highly susceptible to antibiotics and leads to defects in secretion of protein toxins. Despite impairment of various transport functions, no growth...
journal_title:Molecular microbiology
pub_type: 杂志文章
doi:10.1111/j.1365-2958.2010.07245.x
更新日期:2010-08-01 00:00:00
abstract::TOL plasmid pWW0 of Pseudomonas putida contains two operons that specify a pathway for the degradation of aromatic hydrocarbons. The upper pathway operon encodes the enzymes for the oxidation of toluene/xylenes to benzoate/toluates, and the metacleavage pathway operon encodes the enzymes for the further oxidation of t...
journal_title:Molecular microbiology
pub_type: 杂志文章,评审
doi:10.1111/j.1365-2958.1993.tb01222.x
更新日期:1993-09-01 00:00:00
abstract::Summary Successful malaria transmission from the mosquito vector to the mammalian host depends crucially on active sporozoite motility. Sporozoite locomotion and host cell invasion are driven by the parasite's own actin/myosin motor. A unique feature of this motor machinery is the presence of very short subpellicular ...
journal_title:Molecular microbiology
pub_type: 杂志文章
doi:10.1111/j.1365-2958.2009.06828.x
更新日期:2009-12-01 00:00:00