Soybean 90kDa heat shock protein family encoding gene and application thereof

A heat shock protein and encoding gene technology, which is applied to the soybean 90kDa heat shock protein family encoding gene and its application field, can solve the problems of not improving plant stress tolerance, failing to improve Arabidopsis stress tolerance, and few reports on its functions.

Inactive Publication Date: 2013-09-18
NANJING AGRICULTURAL UNIVERSITY
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Problems solved by technology

In plants, although HSP90 has been cloned in several species such as Arabidopsis, maize, rice and wheat, their functions in abiotic stress are rarely reported.
The study of HSP90 in Arabidopsis found that they have a certain response to abiotic stress, but overexpression of these genes in Arabidopsis

Method used

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  • Soybean 90kDa heat shock protein family encoding gene and application thereof
  • Soybean 90kDa heat shock protein family encoding gene and application thereof
  • Soybean 90kDa heat shock protein family encoding gene and application thereof

Examples

Experimental program
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Embodiment Construction

[0032] The methods used in the following examples are conventional methods in the art unless otherwise specified.

[0033] 1) Cloning of soybean 90kDa heat shock protein coding gene GmHSP90s

[0034] Using the reported soybean GmHSP90 protein sequence (GmHSP90-1, accession number FJ222390) as the seed sequence, the soybean genome database was searched by bioinformatics methods, and 12 sequences with homology between 44% and 95% were found. Primers, primer sequences are listed in Table 1.

[0035] Table 1GmHSP90s gene cloning primers

[0036]

[0037]

[0038] The GmHSP90s gene was cloned from the leaves of soybean variety Qihuang 22 by RT-PCR method. Soybean leaves were taken, crushed with a mortar, added to a 1.5ml EP tube filled with lysate, shaken thoroughly, and total RNA was extracted (RNAprep pure Plant Kit, Tiangen, Beijing). According to the reverse transcription kit provided by Japan Toyobo Co., Ltd. qPCR RT Kit (TOYOBO, Japan) was used for reverse transcri...

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Abstract

The invention discloses a soybean 90kDa heat shock protein family encoding gene and an application thereof. The soybean 90kDa heat shock protein and an encoding gene GmHSP90 of the soybean 90kDa heat shock protein, disclosed by the invention, comprise 12 members which are respectively placed in cytoplasm, endoplasmic reticulum, chloroplast and mitochondria; plant overexpression vectors 35S-GmHSP90-pEarleyGate103 respectively constructed by five typical genes convert arabidopsis to obtain expressed transgenic arabidopsis as well as mRNA (Messenger Ribose Nucleic Acid) high-level expression; an overexpression transgenic GmHSP90 plant shows higher resistance to heat stress and low-temperature stress, is certainly resistant to both permeation and drought stress, and also has resistance to high-concentration calcium ions. Therefore, such expressed gene can be used as a target gene for being introduced into the plant through overexpression to improve the comprehensive resistance of the transgenic plant.

Description

technical field [0001] The invention belongs to the field of plant genetic engineering and relates to soybean 90kDa heat shock protein family coding gene and application thereof. Background technique [0002] Adversity stress has a great impact on the yield and quality of crops, and plants often face various stresses such as high temperature, low temperature and drought stress during the growth process. To study the function and mechanism of some stress-related proteins and apply them Breeding with genetic engineering technology has become one of the important methods to improve crop stress tolerance. [0003] Heat Shock Protein (HSP), also known as heat shock protein or stress protein, is a group of proteins produced by cells under the stimulation of stressors (such as high temperature). 90kDa heat shock protein (HSP90) is a highly conserved molecular chaperone ubiquitous in prokaryotes and eukaryotes. Its molecular weight range is 86-92kDa, most of HSP90 are localized in...

Claims

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Application Information

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IPC IPC(8): C12N15/29C12N15/82C12N1/21C07K14/415A01H5/00
Inventor 邢邯徐筋燕薛晨晨赵晋铭郭娜薛冬
Owner NANJING AGRICULTURAL UNIVERSITY
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