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Интеллектуальная Система Тематического Исследования НАукометрических данных |
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Идентификация SINE-подобных элементов в геноме Phytophthora infestans и оценка возможности их применения для сравнительного анализа штаммов//Журнал Российского фитопатологического обществастатья
- Авторы: Лаврова О.И., Еланский С.Н.
- Журнал: Журнал российского фитопатологического общества
- Том: 4
- Год издания: 2003
- Первая страница: 39
- Последняя страница: 45
- Аннотация: The tasks of the work were the identification of conservative DNA fragments similar to A - B box of Short Interspersed Nuclear Elements (SINEs) in Phytophthora infestans genome, selection of PCR-primer for these DNA fragments, and comparative analysis of P. infestans and Stachybotrys chartarum isolates from distant regions using this primer. A search for SINE-similar elements was performed with primers to known A and B boxes of SINEs from other organisms. After PCR-amplification DNA fragments of different sizes were identified. Six DNA fragments (45 - 51 bp) were cloned in E. coli and sequenced. All 6 clones had similar 25-nucleotide part that includes B box. This sequence was used to create revSINE (5'-GGGATCGAACCAGAAGTGACTACGG-3') primer. After PCR-amplification of total P. infestans DNA with RevSINE-primer a great number of DNA fragments of different sizes were obtained. The number of fragments decreased with the increasing of the temperature of the primer melting. The temperature 48 С revealing 94 bands after electrophoresis in PAAG was used for comparative analysis. Cluster analysis of PCR- products did not elucidate any groups of isolates based on there geographic origin or host-plant. Only one isolate from the Sakhalin island differed significantly from other strains. The same results were obtained for S. chartarum isolates. Possible explication is that SINEs - mobile elements and can rapidly change there location in genom. This type of PCR-analysis is characterized by very high resolution and it might be more suitable for other types of researches such as analysis of different features of the strains and gene identification than for population explorations.
- Добавил в систему: Еланский Сергей Николаевич