Sea island cotton receptor albuminoid kinase gene promoter and application thereof

A protein kinase gene and promoter technology, applied in the field of sea island cotton receptor-like protein kinase gene promoter, can solve problems such as slow progress

Inactive Publication Date: 2012-03-14
NANJING AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the elucidation of receptor protein kinase-related genes from the perspective of promoter structure, function and mechanism of action has been slow, and there are few reports so far.

Method used

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  • Sea island cotton receptor albuminoid kinase gene promoter and application thereof
  • Sea island cotton receptor albuminoid kinase gene promoter and application thereof
  • Sea island cotton receptor albuminoid kinase gene promoter and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0075] The reaction system is:

[0076]

[0077] The reaction procedure is:

[0078] A: 1min at 94°C;

[0079]94℃ 30sec;

[0080] 37℃ 3min

[0081] Ramp to 70°C at 0.2°C / S.

[0082] After PCR reaction A is completed, take out the PCR tube, add 2.5 μl of SP1 and continue PCR;

[0083] B: 94°C 30sec

[0084] 70℃ for 5.5min,

[0085] 94°C 30sec.

[0086] Perform PCR reaction B for 25 cycles, and then perform PCR reaction C for 8 cycles

[0087] C: 94°C 30sec

[0088] 50℃ 30sec

[0089] 70°C for 5min.

[0090] The second round of SEFA-PCR amplification:

[0091] The PCR reaction system is:

[0092]

[0093] The reaction procedure is:

[0094]

[0095] Apply the above-described experimental method, design groove primers SP1, SP2 and SP3 according to the untranslated regulatory sequence at the 5' end of the cloned receptor analogous protein kinase, and design the 3' end sefa-PCR primer as follows:

[0096] SP1: 5'-GCAAACACTATCGCTGCTTTCTC-3';

[0097] SP...

Embodiment 2

[0103] We predicted two transcription start sites, A at -221 and A at -1664, with analysis scores of 0.91 and 0.99, respectively. There is a TATA-Box (ATATA) at the -247 / -251 position, and a CAAT-Box at the -257 / -261 and -289 / -293 positions, both of which are necessary for the basic promoters of higher plants Expression Regulatory Elements. There are two ABREs at -366 / -371, and -1127 / -1132, one W-Box at -752 / -756, -1276 / -1280, -1575 / -1579, and one at -613 / Position -620 has a cis-element in the promoter region of a xylem-associated gene. At the same time, the promoter region also includes some regions that bind to transcription factors such as MYB, MYC and WRKY, four Root-Motifs, and original regulatory elements related to light response.

[0104] Example 3:

Embodiment 3

[0106] FI 5'-CCGGAATTCTTTTAATTAGGGTAGACAG-3';

[0107] F2 5'-CCGGAATTCACAATAAAGTCCCGATTAC-3';

[0108] F3 5'-CCGGAATTCAGGTTGGTCTGGCTCGGGATA-3';

[0109] R1 5'-GAAGATCTACCATGTTTAGTGTTATAGATTGAG-3';

[0110] The constructed vector was verified by PCR amplification, enzyme digestion and sequencing, and the results showed that the constructed vector was correct.

[0111] Example 4:

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Abstract

The invention discloses a sea island cotton receptor albuminoid kinase gene promoter and an application thereof. The length of the promoter is 3183bp, nucleotide sequences are shown as SEQ ID NO.1. Receptor albuminoid kinase gene encoding protein products are relevant to the signal conduction of plant stress response, the gene promoter is the promoter with special vascular bundles and is expressed through biological or abiological inducement, the expression level of target genes in the vascular bundles by a gene engineering method through the promoter is enhanced, and the resistance of cotton on greensickness and abiotic stress is improved.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a promoter of a receptor-like protein kinase gene of Gossypium japonicus and its application. Background technique [0002] Plant gene regulation is mainly carried out at the transcriptional level, which is regulated by a variety of cis-acting elements and trans-acting elements. The plant gene promoter is an important cis-acting element, which is a DNA sequence located in the upstream region of the 5' end of the structural gene, which can guide the correct combination of the enzyme and the template, activate RNA polymerase, and make it have the form of initiating specific transcription , which determine the direction and efficiency of transcription and the type of RNA polymerase used, is central to the regulation of transcription. According to their mode of action and function, promoters commonly used in plant genetic engineering can be divided into three categories, nam...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/113A01H5/00
Inventor 张天真赵君郭旺珍张朝军
Owner NANJING AGRICULTURAL UNIVERSITY
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