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Right boundary flanking sequence of exogenous event inserting vector for transgenic rape Oxy-235 and its application

A technology of oxy-235 and exogenous insertion, applied in application, genetic engineering, plant gene improvement, etc., can solve undiscovered problems

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 information about the transgenic rape Oxy-235 event exogenous insertion vector flanking sequence and the use of this sequence to establish event-specific qualitative and quantitative PCR (polymerase chain reaction) detection report

Method used

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  • Right boundary flanking sequence of exogenous event inserting vector for transgenic rape Oxy-235 and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0041] 1. PCR amplification of the sequence flanking the right border of the transgenic rapeseed Oxy-235 event exogenously inserted into the vector

[0042] First preheat 20% SDS to 65°C, take 15ml of 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, 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 pre-warmed 20% SDS, mix well, and water bath at 65°C for at least 30 minutes, in between. Gently shake the test tube; after the water bath, quickly place the centrifuge tube on ice, add 3ml of 3M KAc, mix well, and place on ice for 30 minutes; centrifuge at 5000g at 4°C for 5 minutes; 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 for 15min at 4°C, discard the supernatant, w...

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Abstract

The present invention discloses right boundary flanking sequence of exogenous event inserting vector for transgenic rape Oxy-235 and its application, and relates to bioengineering technology. The present invention obtains right boundary flanking sequence, SEQ No. 1, of exogenous event inserting vector for transgenic rape Oxy-235 with transgenic rape Oxy-235 as material, GenomeWalker of Invitrogen co. for constituting genomic library, and primer pair OXYRB-1 and OXYRB-2, and the jointing primer, and through PCR amplification. The present invention designs primers OXYRG and OXYRV and TaqMan probe OXYRP; and establishes the specific quantitative and qualitative detection method of exogenous event for transgenic rape Oxy-235. The present invention is suitable for specific PCR detection of other Oxy-235 events.

Description

technical field [0001] The invention relates to the detection of transgenic rape in the technical field of bioengineering, in particular to the right border flanking sequence of an exogenous insertion vector of a transgenic rape exogenous gene integration event Oxy-235 and its application. Background technique [0002] In recent years, genetically modified crops such as corn, soybeans, cotton, canola and tomato have been approved for cultivation and production in many countries, and some are processed into food, feed or used as food additives. As the ecological safety and edible safety of genetically modified products have been controversial, more than 30 countries and regions have successively implemented the labeling system of genetically modified products. [0003] According to the sequence information of the exogenous gene in the transgenic product, the combination of the exogenous gene in the transgenic construct, and the integration site of the construct in the genome,...

Claims

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Application Information

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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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