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A new PCR‐based approach to a fast search of a wide spectrum of cry genes from Bacillus thuringiensis strains
Author(s) -
Shevelev A. B.,
Lewitin E.,
Novikova S. I.,
Wojciechowska Ya.A.,
Usacheva E. A.,
Chestukhina G. G.,
Stepanov V. M.
Publication year - 1998
Publication title -
iubmb life
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.132
H-Index - 113
eISSN - 1521-6551
pISSN - 1521-6543
DOI - 10.1080/15216549800203482
Subject(s) - bacillus thuringiensis , gene , genome , genomic dna , biology , accession number (library science) , genetics , strain (injury) , dna , microbiology and biotechnology , genbank , bacteria , anatomy
A pair of highly degenerated primers was adapted to carry out a single‐step PCR‐detection of any known and probably unknown cry genes of classes cry1, cry4 and cry9 encoding for 130 kDa protein δ‐endotoxins in the natural Bacillus thuringiensis i(BT) strains. The Southern hybridization of the product has demonstrated that essentially remote cry‐genes like cry1Aa and cry9A (cryIG) could be represented in the single amplificate if they are simultaneously present in the genome of the analyzed strain. Four genes were detected by the proposed scheme in the iBT ssp. galleriae 11‐67. One of them, gene cry1Ga1 was originally found and cloned using the PCR‐amplification product obtained from the genomic DNA of this strain as a probe. The new gene was completely identical to one cloned by B. Lambert (unpublished, EMBL accession number Z22510) and essentially related to cryIM (EMBL accession number Y09326), renamed according to the new nomenclature as cry1Ga2.