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Plasmid transformation in Bacillus subtilis

✍ Scribed by Iglesias, Antonio ;Ceglowski, Piotr ;Trautner, Thomas A.

Publisher: Springer
Year: 1983
Tongue: English
Weight: 666 KB
Volume: 192
Category: Article
ISSN: 0026-8925
DOI: 10.1007/bf00327660

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✦ Synopsis

Two HindIII generated DNA fragments of 3.0 and 2.3 Kb derived from rRNA genes of B. subtilis were cloned in E. coli with pBR322. The 3.0 Kb fragment could be subcloned in B. subtilis using pC194. However, only the multimeric, but not the monomeric derivatives of this hybrid plasmid were active in transformation of B. subtilis cells. The 2.3 Kb fragment could neither be subcloned in pC194 nor in pPL603, using both cell and protoplast transformation. We attribute the nonclonability of the 2.3 Kb fragment in B. subtilis to the presence of strong promoter activity in this fragment. Direct proof for the presence of a strong promoter, which is apparently responsible for the transcription of the rRNA gene, was obtained in experiments with B. subtilis and E. coli promoter search plasmids.

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