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KMID : 0578319920020020185
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Molecules and Cells
1992 Volume.2 No. 2 p.185 ~ p.190
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Temporal Regulation of 27-kDa Insecticidal Toxin Gene Expression of Bacillus thuringiensis subsp. Israelensis
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Kang, Kyu Younng
Kim, Chang Ryeol/Yun, Han Dae
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Abstract
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A 27-kDa insecticidal toxin gene of Bacillus thuringiensis subsp. israelensis is expressed after onset of sporulation in its own host. However, a cloned 27-kDa toxin gene from B. t. var. monisoni, different by only one base from that of R t. subsp. israelensis [Earp, D. J., and Ellar, D. J. (1987) Nucleic Acids Res. 15, 3619], was expressed in B. subtilis throughout the entire growth phase. To investigate mechanisms how temporal expression of toxin gene is regulated, RNA polymerases of R t subsp. israelensis were purified by polyethylene glycol/dextran phase partitioning and sequential column chromatography on heparin-agarose, DEAF-cellulose and DNA-cellulose. In vitro transcrption and gel retardation assay with a purified RNA polymerase isolated from cells of stationary growth phase indicated that the a subunit of apparent molecular weight 35,000 is involved in transcription of a 27-kDa toxin gene. The sequential biosynthesis of 035-subunit of RNA polymerase, mRNA, and toxin produced by B. t subsp. israelensis provided a correlative evidence that the toxin gene expression is temporally regulated by a subunit of RNA polymerase and its transcriptional specificity of promoter.
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