Bacillus thuringiensis is a gram-positive, spore-forming soil bacterium. It can be used as a biological pesticide, since it produces specific parasporal crystalline inclusions that have proven to be toxic for some widespread insect pests. That is why currently the main aim of the researches concerning these bacteria is creation and examination of new more effective strains to be used for control of the insect pest populations and development of insect resistant plants. The restriction enzymes are widely used for genome analysis of the different strains.
Valicente and da Silva have used restriction enzymes in their study to investigate the genetic diversity within the examined group of Bacillus thuringiensis strains. EcoRI and MseI enzymes were used to create DNA fragments for the further amplification. It was shown that strains 344 and T09 of B. thuringiensis subsp. tolworthi showed the highest similarity of 15%, while the greatest genetic distance of 92% was observed between the strains HD3 Bacillus thuringiensis subsp finitimus and T24 Bacillus thuringiensis subsp neoleonensis.
The purpose of the study of Held et al. was to clone and localize the gene of the protoxin gene of Bacillus thuringiensis subsp. Kurstaki. The restriction enzyme EcoRI was used to digest the DNA molecule and the vector Charon 4A was used for the cloning of the obtained fragments. In the course of the study Held et al. have developed cells that produced protoxin and the investigated gene had chromosomal origin.
Kanda, Tan and Aizawa have integrated the genome of the phage J7W-1 to a plasmid pAF101 of Bacillus thuringiensis strain AFlO. The restriction enzymes EcoRI, HindIII, BglII and PstI were used to digest the plasmid and phage DNA for the further analysis on agarose gel to verify the presence and identify the location of the phage genome. The results have confirmed the successful integration.
Works cited
Held, G. A. et al. 'Cloning And Localization Of The Lepidopteran Protoxin Gene Of Bacillus Thuringiensis Subsp. Kurstaki.'. Proceedings of the National Academy of Sciences 79.19 (1982): 6065-6069. Web.
Kanda, K., Y. Tan, and K. Aizawa. 'A Novel Phage Genome Integrated Into A Plasmid In Bacillus Thuringiensis Strain AF101'. Microbiology 135.11 (1989): 3035-3041. Web.
Valicente, Fernando, and Rosane da Silva. 'Faflp Analysis Of Brazilian Bacillus Thuringiensis Isolates'. SpringerPlus 3.1 (2014): 256. Web.
