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Subcloning and expression of the α-amylase gene from Bacillus subtilis in Xanthomonas campestris

  • Applied Genetics and Regulation
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Summary

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.

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References

  • Atkins DT, Barber CE, Daniels MJ (1987) Transformation of Xanthomonas campestris pathovar campestris with plasmid DNA. J Gen Microbiol 133:2727–2731

    Google Scholar 

  • Balbás P, Soberón X, Merino E, Zurita M, Lomeli FV, Valle F, Flores N, Bolivar F (1986) Plasmid vector pBR322 and its special-purpose derivatives — a review. Gene 50:3–40

    Google Scholar 

  • Barrere GC, Barber CE, Daniels MJ (1986) Molecular cloning of genes involved in the production of the extracellular polysaccharide xanthan by Xanthomonas campestris pv campestris. Int J Biol Macromol 8:372–374

    Google Scholar 

  • Birnboin HC, Doly J (1979) A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res 7:1513–1523

    Google Scholar 

  • Boyer HW, Roulland-Dussoix D (1969) A complementation analysis of the restriction and modification in Escherichia coli. J Mol Biol 41:459–472

    Google Scholar 

  • Bradbury JF (1984) Xanthomonas Dowson 1939 In: Krieg NR, Holt JC (eds) Bergey's manual of systematic bacteriology vol 1. Williams and Wilkins, Baltimore, pp 199–210

    Google Scholar 

  • Caraway WT (1959) A stable starch substrate for the determination of amylase in serum and other body fluids. Am J Clin Path 32:97–99

    Google Scholar 

  • Castro MEB (1988) Análise do gene a α-amylase de Bacillus subtilis visando o desenvolvimento de um vetor bifunctional de expressão-secreção. Master's thesis, University of Brasília, Brasília, Brazil

  • Dragert M, Ehrlich SD (1979) Prolonged incubation in calcium chloride improves the competence of Escherichia coli cells. Gene 6:23–28

    Google Scholar 

  • Daniels MJ, Barber CE, Turner PC, Cleary WG, Sawczyc MK (1984) Isolation of mutants of Xanthomonas campestris pv campestris showing altered pathogenicity. J Gen Microbiol 130:2447–2455

    Google Scholar 

  • Dow JM, Scofield G, Trafford K, Turner PC, Daniels MJ (1987) A gene cluster in Xanthomonas campestris pv campestris required for pathogenicity controls the excretion of polygalacturonate lyase and other enzymes. Physiol Mol Plant Pathol 31:261–271

    Google Scholar 

  • Dye DW, Lelliott RA (1974) Genus II XanthomonasDowson 1939 In: Buchanan RE, Gibbons NE (eds) Bergey's manual of determinative bacteriology. Williams and Wilkins, Baltimore, pp 243–249

    Google Scholar 

  • Figurski H, Helinski DR (1979) Replication of an origin containing derivative of plasmid RK2 dependent on a plasmid function provided in trans. Proc Natl Acad Sci USA 76:1648–1652

    Google Scholar 

  • Fukuda M, Yano K (1985) Construction of broad host range cloning vectors for Gram-negative bacteria. Agric Biol Chem 49:2719–2724

    Google Scholar 

  • Harding E, Cleary JM, Cabanas DK, Rosen AG, Kang K (1987) Genetical and physical analysis of a cluster of genes essential for xanthan gum biosynthesis in Xanthomonas campestris. J Bacteriol 169:2854–2861

    Google Scholar 

  • Haynes WC, Wickerham LJ, Hesseltine CW (1955) Maintenance of cultures of industrially important microorganisms. Appl Microbiol 3:361–368

    Google Scholar 

  • Jeanes A, Pittsley JE, Senti FRJ (1961) Polysaccharide B-1459: a new hydrocolloid polyelectrolyte produced from glucose by bacterial fermentation. J Appl Polym Sci 5:519–520

    Google Scholar 

  • Lanza SA, Rosato YB (1986) Produção de goma xantana por diferentes mutantes e recombinantes de Xanthomonas campestris. Abstracts 13th Reunião Anual Genética de Microorganismos, Ribeirão Preto, Brazil, Jan 29–31, p 84.

  • Maniatis T, Fritsch EF, Sambrook J (1982) Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory Press, New York

    Google Scholar 

  • Murooka Y, Iwamoto H, Hamamoto A, Tatsushi Y (1987) Efficient transformation of phytopathogenic strains of Xanthomonas species. J Bacteriol 169:4406–9

    Google Scholar 

  • Osbourn AE, Barber CE, Daniels MJ (1987) Identification of plant induced genes of the bacterial pathogen Xanthomonas campestris using a promoter-probe plasmid. EMBO J 6:23–28

    Google Scholar 

  • Ramaley RF (1979) Molecular biology of extracellular enzymes. Adv Appl Microbiol 25:37–57

    Google Scholar 

  • Shaw JJ, Kado I (1986) Development of a Vibrio bioluminescence gene set to monitor phytopathogenic bacteria during the ongoing disease process in a non-disruptive manner. Bio/Technology 4:560–564

    Google Scholar 

  • Sjöström M, Wold S, Wieslander A, Rilfors L (1987) Signal peptide amino acid sequences in Escherichia coli contain information related to final protein localization. A multivariate data analysis. EMBO J 6:823–831

    Google Scholar 

  • Souza MBNS (1986) Clonagem molecular e expressão do gene da alfa-amilase be Bacillus sp. Masters' thesis, University of Brasília, Brasília, Brazil

  • Stall RE, Loschke DC, Jones JB (1986) Linkage of copper resistance and avirulance loci on a self-transmissible plasmid in Xanthomonas campestris pv vesicatoria. Phytopathology 76:240–243

    Google Scholar 

  • Stripecke R, Rosato YB (1988) Variabilidade no perfil eletroforético de alfa-esterases e plasmideos em patovares de Xanthomonas campestris. Fitopat Brasil 13:358–361

    Google Scholar 

  • Sumner JB (1925) The determination of sugar in diabetic urine using dinitrosalicylic acid. J Biol Chem 62:287–290

    Google Scholar 

  • Tang JL, Gough CL, Barber CE, Dow JM, Daniels MJ (1987) Molecular cloning of protease gene(s) from Xanthomonas campestris pv campestris: expression in Escherichia coli and role in pathogenicity. Mol Gen Genet 210:443–448

    Google Scholar 

  • Thorne L, Tansey L, Pollock TJ (1987) Clustering of mutations blocking synthesis of xanthan gum by Xanthomonas campestris. J Bacteriol 169:3593–3600

    Google Scholar 

  • Turner P, Barber C, Daniels MJ (1984) Behaviour of the transposons Tn5 and Tn7 in Xanthomonas campestris pv campestris. Mol Gen Genet 195:101–107

    Google Scholar 

  • Williams PH (1980) Black rot: a continuing threat to world crucifers. Plant Dis 64:736–742

    Google Scholar 

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Authors and Affiliations

  1. Department of Genetics and Evolution, Institute of Biology, UNICAMP, 13.081, Campinas, SP, Brazil

    Renata Stripecke & Yoko B. Rosato

  2. Department of Cellular Biology, Institute of Biological Science, University of Brasília, 70.760, Brasília, DF, Brazil

    Spartaco Astolfi-Filho

Authors
  1. Renata Stripecke
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  2. Yoko B. Rosato
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  3. Spartaco Astolfi-Filho
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Stripecke, R., Rosato, Y.B. & Astolfi-Filho, S. Subcloning and expression of the α-amylase gene from Bacillus subtilis in Xanthomonas campestris . Appl Microbiol Biotechnol 31, 512–517 (1989). https://doi.org/10.1007/BF00270786

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  • DOI: https://doi.org/10.1007/BF00270786

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