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Flanking sequence of GhSOCL1 transgenic cotton insertion site and specificity identification method thereof

A technology of insertion site and flanking sequence, which is applied in the field of flanking sequence and specific identification of the insertion site of transgenic GhSOCL1 cotton, and can solve the problems that the flanking sequence of exogenous insertion in transgenic GhSOCL1 cotton has not yet been found in articles and patent reports, etc. To achieve the effect of easy supervision and management

Pending Publication Date: 2021-06-29
INST OF COTTON RES CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, although there have been related patents and literature reports on the flanking sequences of exogenous insertions in transgenic plants, no articles or patent reports related to the flanking sequences of exogenous insertions in transgenic GhSOCL1 cotton have yet been found.

Method used

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  • Flanking sequence of GhSOCL1 transgenic cotton insertion site and specificity identification method thereof
  • Flanking sequence of GhSOCL1 transgenic cotton insertion site and specificity identification method thereof
  • Flanking sequence of GhSOCL1 transgenic cotton insertion site and specificity identification method thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0026] This embodiment provides a flanking sequence of the GhSOCL1 gene cotton insertion site, which includes the left flanking sequence; the nucleotide sequence of the left flanking sequence is as shown in the sequence table SEQ ID NO: 2, and its length is 1348bp; the 1st~912nd site sequence of the left flank sequence is derived from the genome sequence of Zhongmian Institute 24, and the 913th~1348th site sequence of the left flank sequence is derived from the exogenous insertion of GhSOCL1 gene transgenic cotton Fragment sequence. Wherein, the left flank sequence refers to the left border flank sequence of the exogenous insert fragment of the transgenic GhSOCL1 gene cotton.

Embodiment 2

[0028] This embodiment provides a flanking sequence of the GhSOCL1 gene cotton insertion site, which includes the right flanking sequence; the nucleotide sequence of the right flanking sequence is as shown in the sequence table SEQ ID NO:3, and its length is 1269bp; the 1st~466th site sequence of the right flank sequence is derived from the exogenous insert sequence of GhSOCL1 gene transgenic cotton, and the 467th~1269 site sequence of the right flank sequence is derived from the China Cotton Research Institute 24 genome sequence. Wherein, the right flank sequence refers to the right border flank sequence of the exogenous insert fragment of the transgenic GhSOCL1 gene cotton.

[0029] In addition, the acquisition method of the left flank sequence and the right flank sequence provided by the above-mentioned embodiments 1-2 is as follows:

[0030] (1) Extract the genomic DNA of the transgenic GhSOCL1 cotton plant: take the young leaves of the GhSOCL1 transgenic cotton plant and...

Embodiment 3

[0034] This embodiment provides a specific identification primer for the flanking sequence of the insertion site of GhSOCL1 gene transgenic cotton, and the primer is designed according to the left flanking sequence provided in the above-mentioned embodiment 1. Specifically, the primer includes a first primer; the first primer includes a first forward primer designed according to the sequence of the 1st to 912th positions of the left flank sequence and a first forward primer based on the sequence of the 913th to 1348th positions of the left flank sequence The first reverse primer designed by the sequence; wherein, the nucleotide sequence of the first forward primer is as shown in the sequence table SEQ ID NO: 4, which is: 5'-TGCTCTCGATATCGCACACT-3'; the first reverse primer The nucleotide sequence of the primer is shown as SEQ ID NO: 5 in the sequence table, which is: 5'-AGATTGTCGTTTCCCGCCTT-3'.

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Abstract

The invention discloses a flanking sequence of a GhSOCL1 transgenic cotton insertion site and a specificity identification method thereof, and belongs to the technical field of biology. The flanking sequence comprises a left flanking sequence and / or a right flanking sequence; the first to 912th site sequences of the left flanking sequence are derived from a genome sequence of Zhongmiansuo 24, and the 913th to 1348th site sequences of the left flanking sequence are derived from an exogenous insert sequence of transgenic GhSOCL1 cotton; and the first to 466th site sequences of the right flanking sequence are derived from the exogenous insertion element sequence of the GhSOCL1 transgenic cotton, and the 467th to 1269th site sequences of the right flanking sequence are derived from the genome sequence of Zhongmiansuo 24. A specific primer is designed through the flanking sequence, and whether a to-be-detected sample contains a GhSOCL1 transgenic cotton component or not can be judged by judging whether the designed primer and a PCR product of the to-be-detected sample contain a specific amplification band or not.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a flanking sequence of an insertion site of transgenic GhSOCL1 gene cotton and a specific identification method thereof. Background technique [0002] Cotton is one of the most important economic crops in the world, and it is a raw material and an important strategic material for industries such as textiles and fine chemicals. Short season cotton (Short season cotton), as an important cotton variety resource in my country, can well solve the contradiction between grain and cotton for land, and at the same time play an important role in optimizing the agricultural industrial structure. Precociousness is an important target trait in short-season cotton breeding. It is a quantitative trait with complex relationships, which is affected by many factors such as growth period and plant growth and development status. Flowering time directly affects crop maturity, while flower bud differenti...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6895C12N15/11
CPCC12Q1/6895C12Q2600/13
Inventor 马启峰范术丽张晓红乔凯凯张朝军于霁雯喻树迅
Owner INST OF COTTON RES CHINESE ACAD OF AGRI SCI
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