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KMID : 0380020100250020167
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Korean Journal of Biotechnology and Bioengineering
2010 Volume.25 No. 2 p.167 ~ p.172
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Construction of Transformation Method for Streptomyces scabiei ATCC 49173 Producing Phytotoxin
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Jang Bo-Youn
Ha Heon-Su
Choi Sun-Uk
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Abstract
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Streptomyces scabiei producing phytotoxin called thaxtomin, which cause scab disease on economically important crops such as potato. For molecular genetics study of S. scabiei an effective transformation method was established based on conjugal transfer from Escherichia coli ET12567 (pUZ8002) using a phiC31-derived integration vector, pSET152, containing oriT and attP fragments. The high frequency was obtained on MS medium containing 50 mM MgCl2. In addition, the sequence and location of the chromosomal integration attB site of S. scabiei was identified for the first time in the strains producing thaxtomin by the southern blot analysis of exconjugants and the sequencing of plasmid containing DNA flanking the insertion sites from exconjugant chromosome. Similar to the case of Streptomyces species, a single phiC31 attB site of S. scabiei is present within an ORF encoding a pirin-homolog.
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KEYWORD
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Streptomyces scabiei ATCC 49173, transformation, attB site, phytotoxin
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