[The creation of inverted repeats of plant phospholipase С genes fragments]
2006
Pundik, A.N. | Gapeeva, T.A. | Volotovskij, I.D.(Institute of Biophysics and Cell Engineering, Minsk (Belarus))
The scheme for the construction of the inverted repeats of DNA fragments and control of their ligation for the cloning basing on PCR method was suggested. The inverted repeats of tobacco (Nicotiana tabacum) phosphilipase С genes (PI-PLC) were created and inserted into cloning vectors. The presence of the insertion was confirmed by PCR analysis of sudstrates matrix and ligation reaction products. Research results showed that the cloned fragments of the required size were not found by means of restriction and PCR methods after the vectors insertion into bacteria cells. Besides, subsequently the number of the resistant bacteria essentially decreased. Thus, despite the usability of RNA-interference induction in the vegetative cells of chimeric gene containing the inverted repeats, it is necessary to take into consideration the difficulties of cloning and/or detection of corresponding structures which can have the specific character of work with the concrete eukaryotic genes
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