A Rapid Protocol for Screening Genetically Modified Soybeans Based on Positive Plasmid Molecule Containing Multiple Targets
10.13865/j.cnki.cjbmb.2021.10.1329
- Author:
Zong-Yong SHI
1
;
Xuan LIU
1
;
Dong-Mei XU
1
;
Zong-Yong SHI
2
;
Xuan LIU
2
;
Dong-Mei XU
2
;
Yu-Qi ZHANG
2
;
Jian-Hua GAO
2
;
Xia-Ying LI
3
;
Zi-Yan CHEN
3
;
Jin-Gang LIANG
3
;
Hao-Qian WANG
3
;
Xiu-Jie ZHANG
3
;
Hong-Tao WEN
4
;
Jian-Hua GAO
5
Author Information
1. Department of Bioinformatics, College of Life Sciences, Shanxi Agricultural University
2. Crops Ecological Environment Security Inspection and Supervision Center (Taiyuan), Ministry of Agriculture and Rural Affairs
3. Development Center for Science and Technology, Ministry of Agriculture and Rural Affairs
4. Quality and Safety Institute of Agricultural Products, Heilongjiang Academy of Agricultural Sciences
5. Department of Bioscience, College of Life Sciences, Shanxi Agricultural University
- Publication Type:Journal Article
- Keywords:
exogenous elements;
genetically modified soybean;
plasmid;
positive molecule;
screening
- From:
Chinese Journal of Biochemistry and Molecular Biology
2021;37(11):1540-1554
- CountryChina
- Language:Chinese
-
Abstract:
Recently we witness the rising number of genetically modified (GM) soybean (Glycine max) events approved for importing from abroad and developed domestically, so it is urgent to establish a rapid screening protocol that can cover more events with less detection targets and fit the national condition. Additionally, in order to control the detection workload, it is also necessary to construct a multi-targets plasmid (MTP) molecule that can be used as the positive material. In this study, the information of the transgenic elements in 29 GM soybean events was collected and the combinations and frequencies of these elements were analyzed, to establish a novel screening protocol. It includes eight detecting targets, CaMV 35S promoter (P-35S), NOS terminator (T-nos), herbicide tolerance gene pat, E9 terminator (T-E9), insecticidal gene cry1Ac, AHAS promoter (P-AHAS), pin Ⅱ terminator (T-pin Ⅱ), and the event-specific sequence of the transgenic event DP305423, and an endogenous reference gene of soybean Lectin. After validation, the 29 GM soybean events described above can be screened by detection of the nine targets. This is referred to as the “8+1” protocol for GM soybean screening. Then these targeted sequences described in the protocol were simultaneously inserted into a cloning vector to construct the corresponding MTP pDDSC-1910. Finally, we tested whether it could be a positive plasmid. As expected, PCR analysis using pDDSC-1910 as a template showed that specific amplicons were observed with high sensitivity. Therefore, the “8+1” screening protocol for GM soybean was established, and the positive plasmid molecule pDDSC-1910 containing corresponding targets was successfully constructed. These results would facilitate the efficient screening and detection of transgenic soybeans.
