Cloning of sporulation gene spoIVC in Bacillus subtilis |
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Authors: | Masaya Fujita and Yasuo Kobayashi |
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Institution: | (1) Department of Applied Biochemistry, Hiroshima University, 720 Fukuyama, Japan |
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Abstract: | Summary Sporulation gene spoIVC of Bacillus subtilis was cloned by the prophage transformation method in temperate phage 105. The specialized transducing phage, 105spoIVC-1, restored the sporulation of the asporogenous mutant of B. subtilis strain 1S47 (spoIVC133). Transformation experiments showed that the spoIVC gene resides on a 7.3 kb HindIII restriction fragment. Subsequent analysis of the 7.3 kb HindIII fragment with restriction endonuclease EcoRI showed that the spoIVC gene resides on a 3.6 kb EcoRI fragment within the 7.3 kb fragment. The 3.6 kb fragment was recloned into the unique EcoRI site of plasmid pUB110 and deletion derivatives having a deletion within the 3.6 kb insert were constructed. The plasmid carrying the entire spoIVC gene restored the sporulation of strain HU1214 (spoIVC133, recE4) at a frequency of 107 spores/ml, and reduced the sporulation of strain HU1018 (spo
+, recE4) to 107 spores/ml. |
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