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A multipurpose broad host range cloning vector and its use to characterise an extracellular protease gene of Xanthomonas campestris pathovar campestris

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Summary

A multipurpose broad host range plasmid, pIJ3200, was constructed by inserting the polylinker-containing 445 by PvuII fragment of Bluescript M13 into the EcoRI site of the cosmid pLAFR1. Using this vector a protease gene of Xanthomonas campestris pathovar campestris, previously cloned in the recombinant plasmid pIJ3070, was located by deletion to a 2.2 kb DNA region. Sequencing of the protease gene revealed an open reading frame encoding a 580 amino acid polypeptide with molecular weight of 57000. The deduced amino acid sequence showed strong homology with the subtilisin family of serine proteases. This, together with its sensitivity to inhibition by phenylmethylsulphonyl fluoride, suggests that the enzyme belongs to this family of proteases.

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Abbreviations

Rif:

rifampicin

Tc:

tetracycline

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Author notes

  1. Yu-Ning Liu

    Present address: Guangxi Plant Protection Station, Nanning, People's Republic of China

  2. Ji-Liang Tang

    Present address: Laboratory of Molecular Genetics, Guangxi Agricultural College, Nanning, People's Republic of China

Authors and Affiliations

  1. The Sainsbury Laboratory, John Innes Institute, NR4 7UH, Colney Lane, Norwich, UK

    Yu-Ning Liu, Ji-Liang Tang, Belinda R. Clarke, J. Maxwell Dow & Michael J. Daniels

Authors
  1. Yu-Ning Liu
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  2. Ji-Liang Tang
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  3. Belinda R. Clarke
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  4. J. Maxwell Dow
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  5. Michael J. Daniels
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Communicated by H. Hennecke

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Liu, YN., Tang, JL., Clarke, B.R. et al. A multipurpose broad host range cloning vector and its use to characterise an extracellular protease gene of Xanthomonas campestris pathovar campestris . Mol Gen Genet 220, 433–440 (1990). https://doi.org/10.1007/BF00391750

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  • DOI: https://doi.org/10.1007/BF00391750

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