Molecular cloning and expression of Bacillus thuringiensis subsp. galleriae insecticidal crystal protein genes in Escherichia coli. 1989

Q Chen, and Y L Fan
Laboratory of Molecular Biology, Chinese Academy of Agricultural Sciences, Beijing.

The location of the toxin gene of B. thuringiensis subsp. galleriae (H5ab) on the Mr-130Md plasmid is determined by molecular cloning. Double digestion fragments (BamHI and SalI) and PstI restriction fragments as well, from the 130 Md plasmid, of B. thuringiensis subsp. galleriae, are ligated with the cloning vector pAT 153 respectively and transformed into E. coli strain HB 101. Out of 208 transformants, three colonies (FG2, FG9, FG19) give positive hybridization reaction using the HD-1 delta-endotoxin gene as a probe. They are presumed to contain the delta-endotoxin gene of B. thuringiensis subsp. galleriae. Western blot assays indicate that Mr-130 kDal and 68 kDal, crystal proteins produced by clone FG2 react with anticrystal protein antibody. The protein extracts of clone FG2 are lethal to Ostrinia furnacalis (Guenee). This is the first report with regard to the cloning and expression of the B. thuringiensis subsp. galleriae (H5ab) delta-endotoxin gene.

UI MeSH Term Description Entries
D007306 Insecticides Pesticides designed to control insects that are harmful to man. The insects may be directly harmful, as those acting as disease vectors, or indirectly harmful, as destroyers of crops, food products, or textile fabrics. Insecticide
D003001 Cloning, Molecular The insertion of recombinant DNA molecules from prokaryotic and/or eukaryotic sources into a replicating vehicle, such as a plasmid or virus vector, and the introduction of the resultant hybrid molecules into recipient cells without altering the viability of those cells. Molecular Cloning
D004731 Endotoxins Toxins closely associated with the living cytoplasm or cell wall of certain microorganisms, which do not readily diffuse into the culture medium, but are released upon lysis of the cells. Endotoxin
D004926 Escherichia coli A species of gram-negative, facultatively anaerobic, rod-shaped bacteria (GRAM-NEGATIVE FACULTATIVELY ANAEROBIC RODS) commonly found in the lower part of the intestine of warm-blooded animals. It is usually nonpathogenic, but some strains are known to produce DIARRHEA and pyogenic infections. Pathogenic strains (virotypes) are classified by their specific pathogenic mechanisms such as toxins (ENTEROTOXIGENIC ESCHERICHIA COLI), etc. Alkalescens-Dispar Group,Bacillus coli,Bacterium coli,Bacterium coli commune,Diffusely Adherent Escherichia coli,E coli,EAggEC,Enteroaggregative Escherichia coli,Enterococcus coli,Diffusely Adherent E. coli,Enteroaggregative E. coli,Enteroinvasive E. coli,Enteroinvasive Escherichia coli
D005796 Genes A category of nucleic acid sequences that function as units of heredity and which code for the basic instructions for the development, reproduction, and maintenance of organisms. Cistron,Gene,Genetic Materials,Cistrons,Genetic Material,Material, Genetic,Materials, Genetic
D006460 Hemolysin Proteins Proteins from BACTERIA and FUNGI that are soluble enough to be secreted to target ERYTHROCYTES and insert into the membrane to form beta-barrel pores. Biosynthesis may be regulated by HEMOLYSIN FACTORS. Hemolysin,Hemolysins,Hemalysins,Proteins, Hemolysin
D000083722 Bacillus thuringiensis Toxins Endotoxins produced by BACILLUS THURINGIENSIS used in transgenic plants and insecticides. When eaten by a susceptible insect they are protease activated in the insect midgut resulting in death from bacterial septicemia. B thuringiensis Toxins,B. thuringiensis Toxins,Bt Toxin,Bt Toxins,Toxin, Bt,Toxins, B thuringiensis,Toxins, B. thuringiensis
D001413 Bacillus thuringiensis A species of gram-positive bacteria which may be pathogenic for certain insects. It is used for the biological control of the Gypsy moth. Bacilan,Dipel,Thuricide
D001426 Bacterial Proteins Proteins found in any species of bacterium. Bacterial Gene Products,Bacterial Gene Proteins,Gene Products, Bacterial,Bacterial Gene Product,Bacterial Gene Protein,Bacterial Protein,Gene Product, Bacterial,Gene Protein, Bacterial,Gene Proteins, Bacterial,Protein, Bacterial,Proteins, Bacterial
D001427 Bacterial Toxins Toxic substances formed in or elaborated by bacteria; they are usually proteins with high molecular weight and antigenicity; some are used as antibiotics and some to skin test for the presence of or susceptibility to certain diseases. Bacterial Toxin,Toxins, Bacterial,Toxin, Bacterial

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