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Isolation, cloning and expression of DNA fragments reflecting open reading frames of 1.8 and 4.6 kb RNAs of the I-18 C gene of Chironomus tentans

100%

Borowicz B. P.

Journal of Applied Genetics

2005

vol. 46

issue 1

75-83

This study, based on the genetic engineering approach, concerns the regulation of the insect gene expression, but it has also a strong significance for plant protection, which is associated with the future possibility to control insect development more effectively, especially in those species that are pests of cultivated plants. The I-18 C gene of Chironomus tentans is activated by heat-shock and the steroid hormone ? ecdysone. This gene produces different transcripts (at least four) by alternative splicing. Two different open reading frames, i.e. ORFs I and II of the two main transcripts of this gene, i.e. the 1.8 and 4.6 kb RNA, had been isolated, at the DNA level, as the ORFs reflecting DNA fragments, using the polymerase chain reaction, then were cloned into the bluescript vector and finally were cloned into the pET-3a vector to be translated in the T7 RNA polymerase/promoter system of E. coli. As the preliminary outcome of these experiments it was possible to obtain ORF I overexpressed as the polypeptide, whereas ORF II was expressed as the polypeptide that was strongly visible on SDS-polyacrylamide gel. This could facilitate further studies regarding the mechanisms of expression of this gene, including the level of expression of this gene during the C. tentans development.

The use of Bacillus thuringiensis in plant protection: prospects and limitations

88%

Malinowski H.

Biotechnologia

2000

issue 3

81-92

Taking into account the protection of natural environment, it may be expected that the use of Bacillus thuringiensis toxins to control pest insects will be increased. It results from the discovery of novel toxins with different activity spectra and from the creation of more active toxins with the use of genetic engeeniering techniques. Besides, the introduction of transgenic plants such as cotton, potato, tomato, tobacco and others producing the bacterial toxins against more important species of pest insects will significantly reduce the application of chemical insecticides. The possibility of resistance development in insects poses a great threat to the above strategy. The better recognition of biochemical, physiological and genetical mechanisms of resistance to B. thuringiensis toxins will allow to devise a strategy for delaying jnsects resistance. The general principles of this strategy are similar to those used in the case of chemical insecticides and involve the rational application of B. thuringiensis toxins and their rotation with other insecticides.

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Isolation, cloning and expression of DNA fragments reflecting open reading frames of 1.8 and 4.6 kb RNAs of the I-18 C gene of Chironomus tentans

The use of Bacillus thuringiensis in plant protection: prospects and limitations