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Biopolymers and Cell
Article . 2015 . Peer-reviewed
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Biopolymers and Cell
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Detection of stress resistance genes in transgenic maize by multiplex and touchdown polymerase chain reaction

Authors: Bannikova, M.A.;

Detection of stress resistance genes in transgenic maize by multiplex and touchdown polymerase chain reaction

Abstract

????????. ?????????????????? ???????????????? ???????????????? ?????????? ?????????????????? ???? ?????????????????? ???????????????? ?? ?????????????????????? ?????????????????? ???? ???????????? ???????????????????????????? (????????) ???? ?????????????????? (touchdown PCR) ???????????????????????? ???????????????????? ??????????????. ????????????. ?????????????????? ?????????????????? ?????? ???????? ??????????????, ?????????????? ?????? ?????? ???????????? ???? ??????, ???????????????????????? ?????????????????? ?????? ???? ?????????????????? ???????????????????????? ?? ???????????????????? ????????, ?????????????????????? ?????????????????? ??????????????. ????????????????????. ???????????????????? ???????????????? ???????? ???? ?????????????????? ??????, ?????????? ???????????????????? ?????????????????? ???????????????????? ???????????? ?????????????????? ??????, ?????? ???????????????? ?? ?????????????????????????? ?????????????????? ?????????????? ??????????????????, ???? ?????????????????????? ???????? ?????????????????? ???? ?????????????????? ????????????????, ???? ?????????????? ???? ???????????? ???????????????????????????????? ?????????? ?????????????????? BT176, MON810, MON88017, DAS1507, DAS59122, MIR604, GA21, NK603 (????????) ???? Bt11, MON863, MON89034, T25 (???????????????? ??????). ???????? ???? ???????????????? ?????? ???????????????????? ?? ?????????????????????????? ?????????????????????? ??????????????. ????????????????. ???????????????? ???????? ???????? ?????????????????????? ?????? ???????????????? ?????????????? ?????????????? ??????????????????. ???? ?????????????????? ???????????????????????????? ???? ?????????????????? ?????? ?????????? ??????????????, ????????????????????, ???????????? ???? ???????????? ???????????? ???????????????? ???????? ??????????????????????????/?????????????????? ???? ???????????????????? ?? ???????? ?????????????????? ???? ?????????? ??????????????????.

????????. ?????????????????????? ???????????????? ???????????????? ?????????? ???????????????????????? ?? ???????????????????? ???????????????? ?? ?????????????????????? ???????????????? ???? ???????????? ???????????????????????????? (????????) ?? ???????????????????? (touchdown PCR) ???????????????????????? ???????????? ??????????????. ????????????. ?????????????????? ?????????? ?????? ???????? ??????????????, ?????????????? ?????? ???? ???????????? ?? ??????, ???????????????????????? ?????????? ?????? ?? ?????????????????? ???????????????????????? ?? ?????????????????? ????????, ???????????????????????? ???????????? ??????????????. ????????????????????. ?????????????????????? ???????????????? ???????? ?? ???????????????????? ??????, ?????????????????????? ???????????????????????? ?????????????????? ???????????????? ?????????? ??????, ???????????????????? ???? ?????????????????????? ???????????????????????? ???????????????? ????????????????, ?? ?????????????????????????? ???????? ???????????????????????? ?? ???????????????????? ????????????????, ???????????????? ?? ???????????? ?????????????????????????????????? ?????????????? ???????????????? BT176, MON810, MON88017, DAS1507, DAS59122, MIR604, GA21, NK603 (????????) ?? Bt11, MON863, MON89034, T25 (???????????????????? ??????). ???????? ?? ???????????????????? ?????? ?????????????????????? ?? ???????????????????????????? ?????????????????????? ????????????????. ????????????. ???????????????? ?????????????????? ?????? ?????????????????? ?????????????? ???????????????? ????????????????. ?? ?????????????? ???????????????????????????? ?? ???????????????????? ?????? ?????????? ??????????????, ????????????????????, ???????????? ?? ???????????????????? ???????????? ???????????????? ???????? ??????????????????????????/???????????????????????? ?? ???????????????????? ?? ???????? ???????????????????????? ?? ?????????????????? ????????????????????.

Aim. To develop a methodology for detection of the genes of resistance to the stress factors in transgenic maize by multiplex (mPCR) and touchdown polymerase chain reactions. Methods. isolation of total DNA by CTAB method, purification of DNA from RNA and proteins, electrophoresis of total DNA and amplification products in agarose gel, polymerase chain reaction. Results. The protocol of multiplex and touchdown polymerase chain reactions has been developed for simultaneous verification of the quality of total DNA extracted from the studied maize plant samples and detection of the following genes that determine resistance to the stress factors in the transgenic maize and maize transformation events: BT176, MON810, MON88017, DAS1507, DAS59122, MIR604, GA21, NK603 (mPCR), Bt11, MON863, MON89034, T25 (touchdown PCR). The multiplex PCR and touchdown PCR were developed using the reference samples. Conclusions. The proposed protocol of mPCR and touchdown PCR reactions can be used for mass analysis of maize samples to detect the genes of tolerance/resistance to herbicides and genes of resistance to insects reliably, authentically, quickly and cheaply.

Keywords

Molecular and Cell Biotechnologies

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selected citations
These citations are derived from selected sources.
This is an alternative to the "Influence" indicator, which also reflects the overall/total impact of an article in the research community at large, based on the underlying citation network (diachronically).
BIP!Citations provided by BIP!
popularity
This indicator reflects the "current" impact/attention (the "hype") of an article in the research community at large, based on the underlying citation network.
BIP!Popularity provided by BIP!
influence
This indicator reflects the overall/total impact of an article in the research community at large, based on the underlying citation network (diachronically).
BIP!Influence provided by BIP!
impulse
This indicator reflects the initial momentum of an article directly after its publication, based on the underlying citation network.
BIP!Impulse provided by BIP!
0
Average
Average
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gold