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Cotton a kind of avp1 protein and its coding gene and application

A protein and cotton technology, applied in the application, genetic engineering, plant genetic improvement and other directions, can solve problems such as not reaching the industrialization standard, and achieve the effect of improving drought resistance and obvious application prospects.

Inactive Publication Date: 2018-04-27
BIOCENTURY TRANSGENE CHINA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although many research institutions have obtained various types of transgenic plants with certain resistance to salt and drought through modern biotechnology, they have not yet reached the standard of industrialization

Method used

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  • Cotton a kind of avp1 protein and its coding gene and application
  • Cotton a kind of avp1 protein and its coding gene and application
  • Cotton a kind of avp1 protein and its coding gene and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Example 1. Cotton SSH library construction under drought stress:

[0022] The specific method is:

[0023] Using PCR-select from Clontech TM The method shown in the cDNA Subtraction Kit constructs a subtractive library by suppression subtractive hybridization method. During the experiment, the mRNA of the leaves of the drought-treated cotton seedlings was used as the sample (tester), and the mRNA of the leaves of the untreated cotton seedlings was used as the control (driver). The specific steps are briefly described as follows:

[0024] (1) Test materials:

[0025] Ji Mian 14 (National Cotton Medium-Term Bank, acquired by China Cotton Research Institute, uniform number: ZM-30270) was sown on sterilized vermiculite, cultivated at 25 °C, photoperiod 16h / 8h, and watered 1 / 2 2MS medium (9.39mM KNO 3 , 0.625mM KH2PO 4 , 10.3mM NH 4 NO 3 , 0.75mM MgSO 4 , 1.5mM CaCl 2 , 50 μM KI, 100 μM H 3 BO 3 , 100 μM nSO 4 , 30 μM ZnSO 4 , 1 μM Na 2 MoO 4 , 0.1 μM CoCl ...

Embodiment 2

[0036] Cloning of embodiment 2, GhAVP1-1 gene

[0037] In the above-mentioned unigene with homologous sequences, the clone Gh-D087 sequence: SEQ ID No: 3:

[0038] 1 ACCAGTATGT TGGCATTTTC ATGGTTGCTT TTGCAATCTT GATTTTCCTT TTCCTTGGCT

[0039] 61 CGGTAGAGGG TTTCAGCACA AAGAGCAGC CTTGCACTTA TGACAAATCT AAGATGTGCA

[0040] 121 AACCTGCTCT TGCCACTGCT ATATTCAGCA CAGTATCTTT CTTGCTCGGT GCTGTCACTT

[0041] 181 CAGTAGTTTC TGGCTTTCTT GGGATGAAAA TTGCTACCTA TGCTAATGCT CGAACCACCC

[0042] 241 TGGAGGCAAG AAAGGGAGTT GGGAAGGCAT TTATTACTGC ATTCAGATCT GGTGCTGTCA

[0043] 301 TGGGCTTTCT TCTTGCTGCA AATGGTCTTT TGGTGCTTTA CATTGCCATC AACTTATTCA

[0044] 361 AGCTCTACTA TGGTGATGAC TGGGGTGGTC TTTTTGAGGC AATCACTGGT TATGGACTTG

[0045] 421 GAGGTTCATC CATGGCGCTT TTTGGAAGAG TTGGTGGAGG CATCTATACA AAAGCTGCTG

[0046] 481 ATGTGGGTGC TGATCTTGTA GGCAAGGTGG AAAGGAACAT TCCTGAGGAT GACCCTAGAA

[0047] 541 ACCCAGCTGT GATTGCTGAC AATGTTGGGG ATAATGTTGG TGATATCGCT GGGATGGGAT

[0048] 601 CTGATCTTTT TGGGTCCTAT GCTGAAT...

Embodiment 3

[0161] Embodiment 3 GhAVP1-1 gene plant expression vector construction

[0162] The construction process of the plant expression vector rd29A-GhAVP1-1-2300 is shown in Figure 1 .

[0163] The plant binary expression vector pCAMBIA2300 (purchased from Beijing Dingguo Changsheng Biotechnology Co., Ltd.) was selected as the plant expression vector, and the 35S promoter of the NPTII gene containing double enhancers was replaced with the Pnos promoter to reduce the expression of NPTII protein in plants . The inducible promoter rd29A and Tnos were selected as the promoter and terminator of GhAVP1-1 gene. The specific steps are briefly described as follows:

[0164] SEQ ID NO: 11 and SEQ ID NO: 12 use the plant expression vector PBI121 (purchased from Beijing Huaxia Ocean Technology Co., Ltd.) as a template to amplify Pnos, and use TaKaRa's PrimeSTAR HS DNA polymerase. 50 μl PCR reaction system: 10 μl 5×PS Buffer, 3 μl 2.5 mM dNTP, 1.0 μl PBI121, 1.0 μl PrimeSTAR, 10 μM primers SE...

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Abstract

The present invention provides an AVP1 protein derived from cotton and an application thereof in cultivating transgenic plants with improved drought resistance and salt tolerance.

Description

technical field [0001] The invention relates to plant protein and its encoding gene and application, in particular to an AVP1 protein derived from cotton and its application in cultivating transgenic plants with improved drought resistance and salt tolerance. Background technique [0002] Drought restricts the cultivation of crops on more than 40% of the earth's land area, and poses a serious threat to global agricultural production and food supply. Drought is one of the main environmental constraints on crop yield. [0003] The area of ​​saline-alkali soil in the world is very large, about 400 million hectares, accounting for 1 / 3 of irrigated farmland. In semi-arid and arid areas with dry climate, due to low rainfall and intense evaporation, salt accumulates continuously; in coastal areas, soil salt content increases due to seawater intrusion. Saline-alkali soil in my country is mainly distributed in Northwest, North China, Northeast China and coastal regions. With the inc...

Claims

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Application Information

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IPC IPC(8): C07K14/415C12N15/29C12N15/63C12N5/14C12N15/82A01H5/00A01H6/60
CPCC07K14/415C12N15/8273
Inventor 王建胜何云蔚崔洪志
Owner BIOCENTURY TRANSGENE CHINA
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