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Chrysanthemum stress-resistance transcription factor CmMYB2 as well as plant expression vector, construction method and application thereof

A technology of anti-retro-transcription factors and plant expression vectors, applied in the field of molecular biology, can solve the problems of lagging anti-retro-transcription factors and restricting the stress-resistant breeding process of chrysanthemums, and achieve the goal of improving plant stress resistance and salt tolerance Effect

Inactive Publication Date: 2011-08-24
NANJING AGRICULTURAL UNIVERSITY
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AI Technical Summary

Problems solved by technology

Chrysanthemum antiretroviral factors are limited to DREB Cloning and expression characteristics of transcription factors [9,10] , that is, the excavation of key anti-retroviral factors of chrysanthemum is seriously lagging behind, which largely restricts the process of chrysanthemum stress-resistant breeding

Method used

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  • Chrysanthemum stress-resistance transcription factor CmMYB2 as well as plant expression vector, construction method and application thereof
  • Chrysanthemum stress-resistance transcription factor CmMYB2 as well as plant expression vector, construction method and application thereof
  • Chrysanthemum stress-resistance transcription factor CmMYB2 as well as plant expression vector, construction method and application thereof

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Embodiment Construction

[0050] Plant expression vector pCAMBIA1301- CmMYB2 build

[0051] 1. CmMYB2 A clone of:

[0052] Choose chrysanthemum 'Zhongshan purple osmanthus' ( Chrysanthemum grandiflorum ) as material, take 0.05g of young leaves, and extract the total RNA of the leaves according to the instructions of the Trizol RNA Extraction Kit (TaKaRa), and reverse transcribe 1 μg of total RNA into cDNA according to the M-MLV Reverse Transcription Kit (TaKaRa) , digest the cDNA product with RNase, refer to Arabidopsis MYB Sequence information, analyzed by Oligo6.0 software to design primers for amplification CmMYB2 ;

[0053] Upstream primer CmMYB2-F: 5′-ATGGCTACTGTTTCGAAA-3′

[0054] Downstream primer CmMYB2-R: 5′-CTAGTCTATTTTACTAAATCCC-3′

[0055] Using the extracted leaf cDNA as a template, carry out PCR reaction,

[0056] 50 μL reaction system: 5.0 μL of 10×RCR Buffer, 1.0 μL of CmMYB2-F and CmMYB2-R primers (20 μmol L -1 ), dNTP mix 4.0 μL (2.5 mmol·L -1 ), Taq DNA Polymerase 0.2...

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Abstract

The invention discloses a chrysanthemum stress-resistance transcription factor CmMYB2 as well as a plant expression vector, a construction method and an application thereof, belonging to the field of molecular biology. The expression vector pCAMBIA1301-CmMYB2 is constructed with the construction method which comprises the following steps: inserting the CmMYB2 gene into a colon vector pMD19-Tsimplevector (Takara); and after carrying out enzyme digestion on the CmMYB2 gene by BamHI (Takara) and SacI (Takara), connecting to the BamHI locus and SacI locus of an expression vector pCAMBIA1301. The plant expression vector is used for the genetic transformation of plants, the CmMYB2 gene is subjected to over expression under the starting of CaMV35S to synthesize a great quantity of MYB2 proteins, regulate the expression of a downstream target gene and improve plant stress resistance.

Description

technical field [0001] The invention belongs to the field of molecular biology, and relates to chrysanthemum anti-reverse transcription factor CmMYB2 and its plant expression vector, construction method and application. Background technique [0002] The study of transcription factors in the process of plant transcription regulation is a research hotspot in current molecular biology. MYB transcription factor is the largest transcription factor family in plants, members of this family play an important role in regulating a series of important physiological processes in plants, for example, regulating plant second metabolism, cell morphogenesis, controlling development, determining cell fate and Traits and responses of plants to hormones and external environmental factors [1~8] . [0003] chrysanthemum( Chrysanthemum morifolium ) is one of the top ten traditional famous flowers in my country and one of the four major cut flowers in the world. Abiotic stresses such as hig...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/415C12N15/29C12N15/82C12N15/66A01H5/00
Inventor 陈发棣单红陈素梅陈煜刘兆磊宋爱萍房伟民顾春笋
Owner NANJING AGRICULTURAL UNIVERSITY
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