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A transgenic rapeseed sd-rapeseed that aggregates commonly used screening targets and its application

A transgenic and rapeseed technology, applied in application, genetic engineering, plant genetic improvement and other directions, can solve the problems of differences in DNA extraction efficiency of transgenic components, error in detection sensitivity judgment, difficulty in accurately reflecting sensitivity, etc., to reduce labor costs and economic costs, Cost and labor saving, easy application effect

Active Publication Date: 2020-09-18
OIL CROPS RES INST CHINESE ACAD OF AGRI SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, there are many problems in transgenic screening using a mixture of standard substances for specific detection of transformation events: 1) There are no corresponding standard substances for transgenic elements such as the hygromycin resistance gene HPT commonly used in genetic engineering research; 2) Some screening elements It occurs repeatedly in different transformation events, resulting in inconsistent copy numbers of different elements in the positive control, which makes it difficult to accurately reflect the sensitivity of detection; 3) There are differences in the DNA extraction efficiency of different transgenic components, resulting in errors in the judgment of detection sensitivity
[0006] After searching the existing patents and other documents, no reports on the development and application of general reference materials for the screening and detection of transgenic rapeseed have been found.

Method used

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  • A transgenic rapeseed sd-rapeseed that aggregates commonly used screening targets and its application
  • A transgenic rapeseed sd-rapeseed that aggregates commonly used screening targets and its application
  • A transgenic rapeseed sd-rapeseed that aggregates commonly used screening targets and its application

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Embodiment Construction

[0048] Below in conjunction with accompanying drawing and embodiment the present invention is described in detail:

[0049] 1. Obtain T0 transgenic rapeseed through genetic transformation

[0050] 1.1, T-DNA vector construction

[0051] The 11 elements are proposed to include P-35S promoter, P-FMV35S promoter, P-NOS promoter, Bar gene, NPTII gene, HPT gene, Pmi gene, T-NOS terminator, T-35S terminator, T- The g7 terminator and T-e9 terminator were cloned into the binary vector pBI121. The T-DNA region of the pBI121 vector contains a reading frame of the NPTII gene, which is regulated by the P-NOS promoter and the P-NOS terminator, so only the DNA sequences of the remaining 8 elements need to be cloned. Cloning primers were designed based on the nucleotide sequences of the eight screening elements. Using the designed primers, the extracted genomic DNA was used as a template for PCR amplification. The PCR amplification system (50μl) contains: 1×KOD Plus buffer, MgSO4 1mmol / L...

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Abstract

The invention discloses transgenosis oilseed rape SD-rapeseed capable of polymerizing common screening targets and application of the transgenosis oilseed rape SD-rapeseed, and relates to the field of biotechnology including genetic transformation and transgenic test. In the invention, eleven common transgenosis screening elements are guided into double No. 6 in conventional rapeseed varieties through an agrobacterium-mediated method, so that the transgenosis oilseed rape SD-rapeseed is obtained. The fusion sequence of the eleven inserted transgenosis screening elements is shown as SEQ NO.1. The transgenosis oilseed rape SD-rapeseed is taken as a raw material, so that a transgenosis oilseed rape screening detection general base standard substance and a genome DNA standard substance can be prepared. The transgenosis oilseed rape SD-rapeseed is applicable to transgenosis oilseed rape screening detection and preparation of screening detection standard substances, and overcomes the defect that the pollution is easy to pollute when plasmid standard substances are prepared.

Description

technical field [0001] The present invention relates to the field of biotechnology, in particular to a transgenic rapeseed SD-rapeseed that aggregates commonly used screening targets and its application, specifically, a transgenic rapeseed that aggregates 11 commonly used screening targets and its application in transgenic standards obtained through genetic transformation. Applications in substance preparation and transgenic detection. Background technique [0002] Since the large-scale commercialization of genetically modified crops, the safety of genetically modified organisms has been concerned by all walks of life. For this reason, more than 50 countries and regions, including China, the European Union, Japan and South Korea, have promulgated and implemented the safety management of genetically modified organisms. Laws and regulations; in order to protect consumers' right to know, the labeling system has been implemented for genetically modified products, and the labelin...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/84A01H5/00A01H6/20C12N15/10C12Q1/6895
CPCC12N15/8205C12N15/8241C12Q1/6806C12Q1/6895C12Q2600/166
Inventor 武玉花李俊李允静曾新华刘芳吴刚
Owner OIL CROPS RES INST CHINESE ACAD OF AGRI SCI
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