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Nucleic Acids Research, 1983, Vol. 11, No. 12 3973-3987
© 1983

MOLECULAR BIOLOGY

In vitro transcription of the cloned chromosomal crystal gene from Bacillus thuringiensis

André Klier*, Claude Parsot+ and Georges Rapoport*

*Unité de Biochimie Microbienne, Institut Pasteur 28, rue du Docteur Roux, 75724 Paris Cedex 15, France +Unité de Biochimie Cellulaire, Institut Pasteur 28, rue du Docteur Roux, 75724 Paris Cedex 15, France

Received April 7, 1983. Revised May 20, 1983. Accepted May 20, 1983.

We have determined the conditions required for in vitro transcription of the cloned chromosomal crystal gene from Bacillus thuringiensis using either the homologous vegetative RNA polymerase or a sporulation specific form of this enzyme. The gene is actively transcribed by the latter enzyme (form II) but not by the vegetative one. Evidence for a specific recognition between the form II enzyme and the promoter site of the crystal gene was obtained by binding experiments. They showed that the binding is increased by the presence of some additional factors, which change the specificity of the vegetative core-enzyme. The sequence of the promoter has been determined and the startpoint of the transcription deduced. Two hexanucleotide sequences, TACAAT and CCTACG, centered at - 10 and - 35 bp are present, but are somewhat different from the consensus sequences previously described in other bacilli.


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