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Positive plasmid molecule pYMSC-1905 for screening unauthorized transgenic maize

A technology of pymsc-1905 and transgenic corn, which is applied in the direction of using vectors to introduce foreign genetic material, DNA/RNA fragments, microbe determination/inspection, etc. It can solve the problems of lack of standard substances or standard samples, backward detection methods, etc.

Inactive Publication Date: 2020-10-27
黑龙江省农业科学院农产品质量安全研究所 +1
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  • Abstract
  • Description
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  • Application Information

AI Technical Summary

Problems solved by technology

[0004] After more than ten years of construction of the genetically modified detection technology system, my country has established a complete genetically modified corn detection technology standard system, but limited by the supply of raw materials, the development of standard materials or standard samples for daily detection is seriously behind the detection method The development of GMOs often leads to the problem of lack of standard substances or standard samples in the process of genetically modified detection

Method used

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  • Positive plasmid molecule pYMSC-1905 for screening unauthorized transgenic maize
  • Positive plasmid molecule pYMSC-1905 for screening unauthorized transgenic maize
  • Positive plasmid molecule pYMSC-1905 for screening unauthorized transgenic maize

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Example 1 Analysis of transgenic corn materials at home and abroad

[0026] The data of foreign materials is mainly obtained by querying the website of the International Agricultural Biotechnology Application Service Organization (ISAAA) and the Registration Database of Active Modified Organisms (LMO). 229 transformants were found in ISAAA, and 199 transformants were found in LMO. , the transformants listed in the two databases were integrated, and finally 237 transformants (including phytase maize BVLA430101) were sorted out, including 53 independent transformants (Table 1) and 184 compound transformants. Through the analysis, it was found that all the compound transformants were different combinations of 53 independent transformants, so the compound transformants containing this trait could be screened out by detecting the independent transformants.

[0027] Table 1 53 independent transformants in foreign databases

[0028] 32138 Bt10 LY038 MON87411 ...

Embodiment 2

[0032] Example 2 Transgenic corn screening scheme

[0033] By analyzing the data in daily testing and consulting relevant databases, the exogenous elements and genes commonly used in transgenic maize lines, as well as the regulatory elements and marker genes contained in commonly used genetic transformation vectors were obtained, and finally 9 exogenous elements / genes were used for the research. Screening detection of transgenic maize: (1) CaMV 35S promoter, (2) NOS terminator, (3) bar gene, (4) pat gene, (5) CP4-EPSPS gene, (6) NPTⅡ gene , (7) cry1A gene, (8) Rice-Actin promoter, (9) phy gene structure-specific fragment. After the screening elements were determined, 36 independent maize transformant materials that could be collected so far were verified (No. 1-36), and 21 transformants with available sequence information (No. 37-57) were analyzed to obtain Determine its degree of conformity with the database information and its applicability. The verification results showed...

Embodiment 3

[0037] Embodiment 3 The design of transgenic maize screening plasmid molecule

[0038] After determining the target screening element, the transgenic maize screening plasmid molecule was designed according to the sequence characteristics of the element and related standards, and in order to meet the needs of high-throughput detection, this protocol includes both ordinary PCR and fluorescent PCR detection methods (Table 4).

[0039] The determined 9 elements and genes were constructed on the pUC18 vector, plus the standard gene zSSIIb in maize, a total of 10 elements and genes constituted the plasmid standard molecule for screening ( figure 1 ), the plasmid molecule was named pYMSC-1905.

[0040] The selection of the target sequence of each element strives to have the widest applicability, the principle is: (1) the plasmid molecule can be used as a positive control for ordinary qualitative PCR detection, (2) the plasmid molecule can be used as a real-time fluorescent PCR detec...

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Abstract

The invention discloses a positive plasmid molecule pYMSC-1905 for screening unauthorized transgenic maize. The plasmid molecule pYMSC-1905 is obtained by constructing structure specific fragments ofa CaMV 35S promoter, an NOS terminator, a bar gene, a pat gene, a CP4-EPSPS gene, an NPT II gene, a cry1A gene, a Rice-Actin promoter and a phy gene in the transgenic maize and a standard gene zSSIIbin the maize to a plasmid vector pUC18. A plasmid disclosed by the invention can screen out 261 kinds of transgenic maize including complex characters, and covers 91.6% of 285 kinds of maize transformants, and the plasmid molecule can be used as a positive control for common qualitative PCR detection and can also be used as a positive control for real-time fluorescent PCR detection.

Description

technical field [0001] The invention belongs to the technical field of transgene detection, and in particular relates to a non-authorized transgenic maize screening positive plasmid molecule pYMSC-1905. Background technique [0002] Unauthorized genetically modified crops refer to genetically modified crops that have been released into the market without approval, including strains that have been approved in other countries but have not obtained a genetically modified safety certificate in my country, strains that are illegally planted in the field, and strains that are only approved for import as raw materials for processing. strains, and transgenic strains that are still in the experimental stage. [0003] Corn is an important food and feed crop in China. In my country, about 80% of corn is used in the feed industry every year. As one of the four major genetically modified crops, genetically modified corn has reached 59.7 million hectares in the world in 2018. Among them, t...

Claims

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Application Information

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IPC IPC(8): C12N15/63C12N15/11C12Q1/6895
CPCC12N15/63C12Q1/6895C12Q2600/166
Inventor 温洪涛李夏莹杨洋陈子言丁一佳张秀杰关海涛张晓磊张瑞英
Owner 黑龙江省农业科学院农产品质量安全研究所
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