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Multibranch tamarix chinensis dehydrated inducing protein RDZZ gene

A technology for inducing proteins and genes, applied in the field of genetic engineering, can solve problems such as strong salt resistance of salt resistance-related genes, limited gene types and quantities of drought-resistant plants, and lack of genes

Inactive Publication Date: 2005-06-08
NORTHEAST FORESTRY UNIVERSITY
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AI Technical Summary

Problems solved by technology

[0004] At present, in genetic engineering technology, the gene types and quantities of drought-resistant plants are very limited, and genes cloned from highly drought-resistant woody plants are lacking; since most drought-resistant genes are cloned from non-stress-resistant plants, such as Arabidopsis thaliana), rice (Oryza sativa) and other plants, and the salt-resistance-related genes of non-drought-resistant plants obviously cannot have strong salt-resistance; moreover, most of the drought-resistance-related genes come from herbaceous plants, microorganisms, etc., because they Genetic differences from woody plants where these genes may not be as effective

Method used

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Examples

Experimental program
Comparison scheme
Effect test

specific Embodiment approach 1

[0006] Specific implementation mode one: this implementation mode is realized in this way:

[0007] (1) Extract the total RNA of Tamarix:

[0008] a. Add CTAB (cetyltrimethylammonium bromide) extraction buffer (2% (w / v) CTAB, NaCl 1.4mol / L, sodium tetraborate 0.025mol / L, pH=9 to the centrifuge tube ) and 10% (V / V) β-mercaptoethanol to obtain the extract, and preheat it in a water bath at 65°C for 5 minutes;

[0009] b. Cool and grind the Tamarix material with liquid nitrogen, add it to the extract, mix well, and put it in a water bath at 65°C for 10 minutes;

[0010] c. Add an equal volume of Tris phenol / chloroform mixture, centrifuge at 12000g for 5 minutes, take the supernatant, and repeat the phenol / chloroform extraction once;

[0011] d. Add an equal volume of chloroform to the supernatant, centrifuge at 12,000 g for 5 minutes, take the supernatant, add LiCl with a final concentration of 2 mol / L, and place in an ice bath for 3 hours;

[0012] e. Centrifuge at 15000g at ...

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Abstract

A dehydration inducing protein RD22 gene of the branchy tamarisk which has strong resistance to salt is a cloned respondent gene to salt and drought stress. Its coded region has the 1170 bp of length and 389 coded amino acids. Its sequence is obviously different from that of the dehydration inducing protein gene for other varieties. Its expression after the drought stress is increased by more than 3 times, so it is a gene associated with drought resistance.

Description

Technical field: [0001] The invention belongs to the technical field of genetic engineering and relates to a dehydration-inducing protein gene of tamarisk willow. Background technique: [0002] At present, the application of transgenic technology to improve plant resistance has become a consensus, and there have been reports of various genes being patented. Tamarix is ​​extremely drought- and salt-tolerant, indicating that there are a series of genes with strong stress resistance in its body. Therefore, it is a good material for carrying out resistance genes. [0003] The gene cloning technology is to determine the target gene to be cloned, and then obtain the fragment or full length of the desired gene by constructing a cDNA library and EST analysis, RT-PCR, yeast two-hybrid and other technologies, and apply the RACE method to the gene fragment Or cDNA library screening method to obtain the full-length gene. [0004] At present, in genetic engineering technology, the gen...

Claims

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Application Information

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IPC IPC(8): C12N15/29
Inventor 杨传平刘桂丰王玉成姜静
Owner NORTHEAST FORESTRY UNIVERSITY
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