Effect Of Transposon Tn5 On Exopolysaccharide Production By Xanthomonas Campestris

[No abstract available]14359960

Regulation Of The Htpx Gene Of Xylella Fastidiosa And Its Expression In E. Coli

Xylella fastidiosa was the first phytopathogen to be completely sequenced, and its genome revealed several interesting features to be used in functional studies. In the present work, the htpX gene, which encodes a protein involved in the heat shock response in other bacteria, was analyzed by RT-PCR by using cells derived from different cultural conditions. This gene was induced after a temperature upshift to 37°C after growth in minimal medium, XDM, but showed constitutive expression in rich medium or in XDM plus plant extracts. Sequences upstream to the htpX gene, containing a putative regulatory region, were also transferred to E. coli, and the thermoregulation was maintained in the new host, since it was constitutively transcribed at 37°C or 45°C in all culture media tested, but not at 28°C in minimal culture medium. The gene was also cloned into the expression vector pET32Xa/LIC, and the expression of the corresponding protein was confirmed by Western blotting.486391395Beretta, M.J.G., Harakava, R., Chagas, C.M., First report of Xylella fastidiosa in coffee (1996) Plant Dis, 80, p. 821Bhattacharyya, A., Stilwagen, S., Reznik, G., Draft sequencing and comparative genomics of Xylella fastidiosa strains reveal novel biological insights (2002) Genome Res, 12, pp. 1556-1563Chang, C.J., Garnier, M., Zreik, L., Rossetti, V., Bove, J.M., Culture and serological detection of the xylem-limited bacterium causing citrus variegated chlorosis and its identification as a strain of Xylella fastidiosa (1993) Curr Microbiol, 27, pp. 137-142Costa De Oliveira, R.C., Yanai, G., Muto, N.H., Leite, D.B., Souza, A.A., Coletta-Filho, H.D., Machado, M.A., Nunes, L.R., Competitive hybridization on spotted microarrays as a tool to conduct comparative genomic analysis of Xylella fastidiosa strains (2002) FEMS Microbiol Lett, 216, pp. 15-21Davis, M.J., French, W.J., Schaad, N.W., Axenic culture of the bacteria associated with phony disease of peach and plum leaf scald (1981) Curr Microbiol, 6, pp. 309-314Dow, J.M., Daniels, M.J., Xylella genomics and bacterial pathogenicity to plants (2000) Yeast, 17, pp. 263-271Hopkins, D.L., Xylella fastidiosa - Xylem-limited bacterial pathogen of plants (1989) Annu Rev Phytopathol, 27, pp. 271-290Hopkins, D.L., Purcell, A.H., Xylella fastidiosa: Cause of Pierce's disease of grapevine and other emergent diseases (2002) Plant Dis, 86, pp. 1056-1066Jenkins, D.E., Auger, E.A., Matin, A., Role of RpoH, a heat shock regulator protein, in Escherichia coli carbon starvation protein synthesis and survival (1991) J Bacteriol, 173, pp. 1992-1996Kornitzer, D., Teff, D., Altuvia, S., Oppenheim, A.B., Isolation, characterization, and sequence of an Escherichia coli heat-shock gene, htpX (1991) J Bacteriol, 173, pp. 2944-2953Kyte, J., Doolittle, R.F., A simple method for displaying the hydropathic character of a protein (1982) J Mol Biol, 157, pp. 105-132Lemos, E.G.M., Alves, L.M.C., Campanharo, J.C., Genomics-based design of defined growth media for the plant pathogen Xylella fastidiosa (2003) FEMS Microbiol Lett, 219, pp. 39-45Mehta, A., Rosato, Y.B., Differentially expressed proteins in the interaction of Xanthomonas axonopodis pv. citri with leaf extract of the host plant (2001) Proteomics, 1, pp. 1111-1118Mircetich, S.M., Lowe, S.K., Moller, W.J., Nyland, G., Etiology of almond leaf scorch disease and transmission of causal agent (1976) Phytopathology, 66, pp. 17-24Nagai, H., Yano, R., Erickson, J.W., Yura, T., Transcriptional regulation of the heat shock regulatory gene rpoH in Escherichia coli: Involvement of a novel catabolite-sensitive promoter (1990) J Bacteriol, 172, pp. 2710-2715Neidhardt, F.C., Vanbogelen, R.A., Lau, E.T., Molecular cloning and expression of a gene that controls the high-temperature regulon of Escherichia coli (1983) J Bacteriol, 153, pp. 597-603Nunes, L.R., Rosato, Y.B., Muto, N.H., Microarray analysis of Xylella fastidiosa provides evidence of coordinated transcription control of laterally transferred elements (2003) Genome Res, 13, pp. 570-578Raivio, T.L., Silhavy, T.J., Periplasmic stress and ECF sigma factors (2001) Annu Rev Microbiol, 55, pp. 591-624Rosato, Y.B., Neto, J.R., Miranda, V.S., Carlos, E.F., Manfio, G.P., Diversity of a Xylella fastidiosa population isolated from Citrus sinensis affected by citrus variegated chlorosis in Brazil (1998) Syst Appl Microbiol, 21, pp. 593-598Rossetti, V., Garnier, M.B., Beretta, J.M., Teixeira, M.J.G., Quaggio, A.R.R., Negri, J.A., Presénce de bactéries dans lê xyléme d'orangers atteints de chorose variegée, une nouvelle maladie dês agrumes au Brésil (1990) Comp Rend Acad Sci Ser, 3, pp. 345-349Sambrook, J., Fritsch, E.F., Maniatis, T., (1989) Molecular Cloning, A Laboratory Manual, 2nd Ed., , Cold Spring Harbor, NY: Cold Spring Harbor Laboratory PressShimohata, N., Chiba, S., Saikawa, N., Ito, K., Akyiama, Y., The Cpx stress response system of Escherichia coli senses plasma membrane proteins and controls HtpX, a membrane protease with a cytosolic active site (2002) Genes Cells, 7, pp. 653-662Simpson, A.J.G., Reinach, F.C., Arruda, P., The genome sequence of the plant pathogen Xylella fastidiosa (2000) Nature, 406, pp. 151-157Wells, J.M., Raju, B.C., Hung, H.Y., Weisburg, W.G., Mandelcopaul, L., Brenner, D.J., Xylella fastidiosa gen-nov, sp-nov, Gram-negative, xylem-limited, fastidious plant bacteria related to Xanthomonas-Spp (1987) Int J Syst Bacteriol, 37, pp. 136-143Yasukawa, T., Kaneiishii, C., Maekawa, T., Fujimoto, J., Yamamoto, T., Ishii, S., Increase of solubility of foreign proteins in Escherichia coli by coproduction of the bacterial thioredoxin (1995) J Biol Chem, 270, pp. 25328-2533

The Role Of A Cloned α-amylase Gene In Fermentation By Xanthomonas Campestris

[No abstract available]13466166

Proteins Induced By Xanthomonas Axonopodis Pv. Passiflorae With Leaf Extract Of The Host Plant (passifiorae Edulis)

Two-dimensional gel electrophoresis was used to identify differentially displayed proteins during treatment of Xanthomonas axonopodis pv. passiflorae in media containing leaf extract of the compatible (passion fruit) and incompatible (tomato) hosts. The results showed that at different times of treatment (5, 25 and 45 h) the global expression of proteins was almost identical in cells grown in minimal medium (MM) and in medium containing leaf extract of the incompatible host (MMT). The protein patterns of cells grown in medium containing passiflorae (MMP) leaf extract and MM were also compared enabling the detection of 17 differential spots. Most of the proteins were induced at earlier times of incubation (5 h) and maintained until 45 h in MMP. By using another carrier ampholyte range, seven additional proteins were identified in MMP treated cells. Five proteins, including one constitutive, two induced and two up-regulated in MMP were microsequenced. All sequences were found in the genome of xanthomonads sharing high level of identity (88-100%). Fructose biphosphate aldolase was expressed in all media employed. A putative membrane-related protein and a hypothetical protein were novel proteins induced specifically by the passiflorae extract. An inorganic pyrophosphatase and a hypothetical protein that showed similarity to the yciF gene of Salmonella thyphimurium were up-regulated in MMP.3195102Vauterin, L., Hoste, B., Kersters, K., Swings, J., (1995) Int. J. Syst. Bacteriol., 45, pp. 472-489Gonçalves, E.R., Rosato, Y.B., (2000) Int. J. Syst. Evol. Microbiol., 50, pp. 811-821Alfano, J.R., Colmer, A., (1997) J. Bacteriol., 79, pp. 5655-5662Jungblut, P., Wittmann-Liebold, B., (1995) J. Biotechnol., 41, pp. 111-120Mehta, A., Rosato, Y.B., (2001) Proteomics, 1, pp. 1111-1118Guerreiro, N., Djordjevic, M.A., Rolfe, B.G., (1999) Electrophoresis, 20, pp. 818-825Natera, S.H., Guerreiro, N., Djordjevic, M.A., (2000) Mol. Plant Microbe Interact., 13, pp. 995-1009Dainese-Hatt, P., Fischer, H.M., Hennecke, H., James, P., (1999) Electrophoresis, 20, pp. 3514-3520Münchbach, M., Dainese, R., Staudenmann, W., Narberhaus, F., James, P., (1999) Eur. J. Biochem., 264, pp. 39-48Guerreiro, N., Redmond, J.W., Rolfe, B.G., Djordjevic, M.A., (1997) Mol. Plant Microbe Interact., 10, pp. 506-516Turner, T., Barber, C., Daniels, M., (1984) Mol. Gen. Genet., 195, pp. 101-107Schulte, R., Bonas, U., (1992) J. Bacteriol., 174, pp. 815-823Vauterin, L., Swings, J., Kersters, K., (1991) J. Gen. Microbiol., 137, pp. 1677-1687Laemmli, U.K., (1970) Nature (Lon.), 227, pp. 680-685De Mot, R., Vanderleyden, J., (1989) Can. J. Microbiol., 35, pp. 960-967O'Farrell, P.H., (1975) J. Biol. Chem., 250, pp. 4007-4021Altschul, S.F., Gish, W., Miller, W., Myers, E.W., Lipman, D.P., (1990) J. Mol. Biol., 215, pp. 403-410Da Silva, A.C.R., Ferro, J.A., Reinach, F.C., Farah, C.S., (2002) Nature, 417, pp. 459-463Sow, P., Demain, A.L., (1979) Appl. Environ. Microbiol., 31, pp. 1186-1192Schmid, K., Schupfner, M., Schmid, R., (1982) J. Bacteriol., 151, pp. 68-76Sprenger, G.A., Lengeler, J.W., (1988) J. Gen. Microbiol., 134, pp. 1635-1644Bogs, J., Geider, K., (2000) J. Bacteriol., 182, pp. 5351-5358Johnson, Bhatnagar, D.R., Knoll, R.S., Gordon, L.J., Annu, J.I., (1994) Rev. Biochem., 63, pp. 869-914Ninchuk, Z., Marois, E., Kjemtrup, S., Leister, R.T., (2000) Cell, 101, pp. 353-363Puri, N., Jenner, C., Bennett, M., Stewart, R., (1997) Mol. Plant-Microbe Interact., 10, pp. 247-256Fang, F.C., Libby, S.J., Buchmeier, N.A., Loewenm, P.C., (1992) Proc. Natl. Acad. Sci. USA., 89, pp. 11978-11982Norel, F., Rosbe-Saule, V., Popoff, M.Y., Coynault, C., (1992) FEMS Microbiol. Lett., 9, pp. 271-276Gulig, R.A., Danbara, H.H., Guiney, D.G., Lax, A.J., (1993) Mol. Microbiol., 7, pp. 825-830Ibanez-Ruiz, M., Robbe-Saule, V., Hermant, D., Labrude, S., Norel, F., (2000) J. Bacteriol., 182, pp. 5749-5756Loewen, P.C., Hu, B., Strutinsky, J., Sparling, R., (1998) Can. J. Microbiol., 44, pp. 707-717Simpson, A., Reinach, F., Arruda, P., Abrev, F.A., (2000) Nature (Lon), 406, pp. 151-157Cooperman, B.S., Alexander, A.B., Lahti, R., (1992) Trends Biochem. Sci., 17, pp. 262-266Sarafian, V., Kim, Y., Poole, R.J., Rea, R.A., (1992) Proc. Natl. Acad. Sci. USA, 89, pp. 1775-177

Subcloning And Expression Of The α-amylase Gene From Bacillus Subtilis In Xanthomonas Campestris

A hybrid plasmid (pAP1) containing the α-amylase gene from Bacillus subtilis was constructed, using the pMFY40 plasmid as a cloning vector. The pAP1 plasmid was introduced into Xanthomonas campestris cells either by conjugation or transformation. The pAP1 plasmid proved to be stable under an antibiotic selection medium. The relative orientation of transcription of the α-amy gene in plasmid pMFY40 was deduced from single and double digestion with restriction enzymes. The expression of the amy gene was detected in non-amylolytic strains of X. campestris and Escherichia coli using an iodine staining assay in solid medium and measuring starch degradation and production of reducing sugars in liquid medium. © 1989 Springer-Verlag.315-651251

Characterization By Polyacrylamide Gel Electrophoresis Of Whole-cell Proteins Of Some Bacillus Thuringiensis Subsp. Israelensis Strains Isolated In Brazil

Of 14 strains of Bacillus thuringiensis selected from 956 isolates from soil samples from Brazil, 12 were toxic to larvae of Aedes fluviatilis and two were nontoxic. Nine of the 14 strains were serotyped as subspecies israelensis (serotype 14), one as subspecies kurstaki (serotype 3a 3b) one as subspecies morrisoni (serotype 8a 8b) and three did not agglutinate any antisera. Electrophoresis of whole cell proteins showed that all subsp. israelensis strains formed a homogeneous group which included two non-typable toxic strains, and could be readily distinguished from reference strains toxic for lepidoptera or coleoptera.17110410

Effect Of Parb On Plasmid Stability And Gene Expression In Xanthomonas Campestris

The stabilization locus parB was subcloned into the broad host range plasmid pAP2, which contains the alpha-amylase gene from Bacillus subtilis, and introduced into Xanthomonas campestris pv campestris and X.c.pv manihotis. Analysis of the stability of plasmid pAP2 (parB-) and pAP23 (parB+) showed that the parB locus decreased significantly the plasmid loss rate mainly by X.c.pv campestris. The lower efficiency of stabilization in X.c.pv manihotis was probably due to the incompatibility system between the native plasmids and the newly introduced pAP23. Although parB had conferred high stability, it determined a lower rate of alpha-amylase activity even by the strain Cm where its stabilization rate was higher.14623323