Flanking sequence of GhFPF1 transgenic cotton insertion site and specificity identification method thereof

A technology of inserting sites and flanking sequences, which is applied in the determination/inspection of microorganisms, biochemical equipment and methods, DNA/RNA fragments, etc., and can solve the problem of foreign insertion of flanking sequences in transgenic GhFPF1 cotton. Articles and patent reports have not yet been found, etc. problem, 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

Although there have been related patents and literature reports on the flanking sequences of exogenous insertions in transgenic plants,

Method used

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  • Flanking sequence of GhFPF1 transgenic cotton insertion site and specificity identification method thereof
  • Flanking sequence of GhFPF1 transgenic cotton insertion site and specificity identification method thereof
  • Flanking sequence of GhFPF1 transgenic cotton insertion site and specificity identification method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] This embodiment provides a flanking sequence of a transgenic GhFPF1 gene cotton insertion site, which includes a left flanking sequence; the nucleotide sequence of the left flanking sequence is shown in the sequence table SEQ ID NO: 2, and its length is 937bp; the sequence of the 1st to 501st sites of the left flank sequence is derived from the genome sequence of China Cotton Research Institute 24, and the sequence of the 502nd to 937th sites of the left flank sequence is derived from the exogenous insertion of transgenic cotton with GhFPF1 gene Fragment sequence. Wherein, the left flank sequence refers to the left border flank sequence of the exogenous insert fragment of the transgenic GhFPF1 cotton.

Embodiment 2

[0027] This embodiment provides a kind of flanking sequence of transgenic GhFPF1 gene cotton insertion site, and it comprises right flanking sequence; The nucleotide sequence of described right flanking sequence is as shown in sequence table SEQ ID NO:3, and its length is 967bp; the 1st~466th site sequence of the right flank sequence is derived from the exogenous insert sequence of GhFPF1 gene transgenic cotton, and the 467th~967th 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 cotton with GhFPF1 gene.

[0028] 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:

[0029] (1) Extract the genomic DNA of the transgenic GhFPF1 cotton plant: take the young leaves of the transgenic GhFPF1 c...

Embodiment 3

[0033] This embodiment provides a specific identification primer for the flanking sequence of the insertion site of transgenic GhFPF1 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 1st to 501st site sequence of the left flank sequence and a 502nd to 937th site according to 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'-GCACCCAGATAATACGGGCT-3'; the first reverse primer The nucleotide sequence of the primer is shown in SEQ ID NO: 5 in the sequence table, which is: 5'-CCTTCAACGTTGCGGTTCTG-3'.

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Abstract

The invention discloses a flanking sequence of a GhFPF1 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 1st-501th site sequences of the left flanking sequence are derived from genome sequences of the CRI 24, and the 502th-937th site sequences of the left flanking sequence are derived from exogenous insert fragment sequences of GhFPF1 transgenic cotton; the 1st-466th site sequences of the right flanking sequence are derived from exogenous insertion fragment sequences of GhFPF1 transgenic cotton, and the 467th-967th site sequences of the right flanking sequence are derived from genome sequences of the CRI 24. According to the invention, a specific primer is designed through the flanking sequence, and whether a to-be-detected sample contains a GhFPF1 transgenic cotton component 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 GhFPF1 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. With the development of biotechnology, transgenic technology is a new way to improve important traits of crops. The advantage of transgenic breeding is that it can realize cross-species gene discovery, broaden the utilization range of genetic resources, and realize the targeted and efficient transfer of known functional genes. Biological acquisition of specific traits needed by humans provides a new technical approach for the cultivation of new varieties of high-yield, high-quality, and high-resistance agricultural organisms. The Cotton Research Institute of th...

Claims

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Application Information

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IPC IPC(8): C12Q1/6895C12Q1/686C12N15/11
CPCC12Q1/686C12Q1/6895C12Q2600/13C12Q2565/125
Inventor 马启峰王小艳范术丽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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