|Table of Contents|

Flanking Sequence Isolation of Salt-Tolerant Transgenic Event FA8015 of Soybean and Quantitative PCR Test
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《大豆科学》[ISSN:1000-9841/CN:23-1227/S]

Issue:
2018年06期
Page:
854-859
Research Field:
Publishing date:

Info

Title:
Flanking Sequence Isolation of Salt-Tolerant Transgenic Event FA8015 of Soybean and Quantitative PCR Test
Author(s):
MA Kuo1; 2;  ZHONG Xiao-fang2;  NIU Lu2;  ZHAO Qian-qian2;  YANG Xiang-dong2
（1 School of Life Science, Jilin Normal University, Siping 136000, China; 2 Agro-Biotechnology Institute, Jilin Academy of Agricultural Sciences / Jilin Provincial Key Laboratory of Agricultural Biotechnology, Changchun 130033, China）
Keywords:
Soybean;  Salt tolerance;  Transgenic event;  Re-sequencing;  Flanking sequence;  Qualitative PCR detection
PACS:
-
DOI:
1011861/jissn1000-98412018060854
Abstract:
In our previous study, we introduced the betaine-aldehyde dehydrogenase encoding gene AhBADH from Atriplex hortensis into the soybean cultivar Williams 82 to generate the salt-tolerant transgenic event FA8015 To advance safety assessment of the transgenic event, in this study, we further analyzed the flanking sequences of the foreign T-DNA insertion and established an event-specific detection method based on these flanking sequences Southern blot analysis confirmed the single-copy insertion of the T-DNA in the genome of the transgenic event FA8015 Genome re-sequencing method was used to obtain the genomic sequences of transgenic event FA8015 The integration site of T-DNA in FA8015 was preliminary determinated compared with Williams 82 genome information Then the primers were designed according to the flanking sequences of integration site for PCR amplification After sequencing these fragments, a 1 160 bp fragment was amplified from left border of T-DNA, including 405 bp fragment from soybean genome and 750 bp sequence from T-DNA, and a 1 254 bp product was obtained from right border with 601 bp T-DNA fragment and 653 bp soybean genome sequence These results indicated that one copy of T-DNA in FA8015 was integrated into soybean genome at 2512882 on Chr15 in accord with Southern blot and genome re-sequencing Event-specific primers were designed base on fragment sequencing and then event-specific PCR was constructed This method could identify this transgenic event quickly and specifically And it also provided support for the safety and supervision of the transgenic event FA8015 and its derivatives

References:

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Last Update: 2018-12-04