Left boundary flanking sequence of exogenous event inserting vector for transgenic rape T45 and its application

A technology of exogenous insertion and flanking sequences, which can be used in applications, genetic engineering, plant genetic improvement, etc., and can solve problems such as specificity and characterization that have not yet been found

Inactive Publication Date: 2007-08-22
INST OF OIL CROPS RES CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0010] After searching the existing patents and other literatures, no report has been found on the transgenic rapeseed T45 event exogenous insertion vector flanking the left border sequence and using this sequence to establish event-specific qualitative and quantitative PCR (polymerase chain reaction) detection

Method used

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  • Left boundary flanking sequence of exogenous event inserting vector for transgenic rape T45 and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0041] 1. PCR amplification of sequences flanking the left border of the transgenic rapeseed T45 event exogenous insertion vector

[0042] First preheat 20% SDS to 65°C, take 15ml SDS extraction buffer (0.1TrisHCl, 0.05MEDTA, 1M NaCl pH8.0) and add it to a 50ml centrifuge tube, then add 2.5μl β-mercaptoethanol and mix well; liquid nitrogen Grind about 3g of leaves, transfer the powder to 50ml of a 50ml centrifuge tube containing extraction buffer, shake and mix on a shaker, add 2ml of preheated 20% SDS, mix well, and bathe in 65°C water for at least 30 minutes, during which time Shake the test tube gently; after the water bath, quickly place the centrifuge tube on ice, add 3ml 3M KAc, mix well, and place it on ice for 30 minutes; centrifuge at 5000g at 4°C for 5min; transfer the supernatant to a new 50ml centrifuge tube, Add 2 / 3 volume of isopropanol, mix well, place at -20°C for more than 30min; centrifuge at 6000g, 4°C for 15min, pour off the supernatant, wash with 75% ethan...

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Abstract

The present invention discloses left boundary flanking sequence of exogenous event inserting vector for transgenic rape T45 and its application. The present invention obtains left boundary flanking sequence, SEQ No. 1, of exogenous event inserting vector for transgenic rape T45 with transgenic rape T45 as material. Of the left boundary flanking sequence, the 1-255 places bases are the same of that of the inserted vector sequence, and the 256-2154 places bases have no homology with that of the inserted vector sequence and are rape genome sequence. The present invention establishes the specific quantitative and qualitative detection method of exogenous event for transgenic rape T45 for the other specific event PCR detection of transgenic rape T45.

Description

technical field [0001] The invention relates to the detection of transgenic rapeseed in the technical field of bioengineering, in particular to an exogenous insertion vector flanking sequence of an exogenous gene integration event T45 of transgenic rapeseed and its application. Background technique [0002] In recent years, genetically modified crops such as corn, soybean, cotton, rapeseed and tomato have been approved to be planted and produced in many countries, and some have been processed into food, feed or used as food additives. Since the ecological safety and food safety of genetically modified products have been controversial, more than 30 countries and regions have successively implemented genetically modified product labeling systems. [0003] According to the sequence information of the exogenous gene in the transgenic product, the combination of the exogenous gene on the transgenic construct, and the integration site of the construct on the genome, gene-specific,...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/82C12N15/29C12N15/65C12Q1/68
Inventor 卢长明吴刚武玉花肖玲
Owner INST OF OIL CROPS RES CHINESE ACAD OF AGRI SCI
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