Rubber plant translationally controlled tumor protein, encoding gene thereof and application of rubber plant translationally controlled tumor protein

A protein and gene technology, applied in the rubber tree translation control tumor protein and its encoding gene and application field, can solve the problems of dead skin, latex coagulation, blocked milk ducts, etc.

Active Publication Date: 2012-11-21
RUBBER RES INST CHINESE ACADEMY OF TROPICAL AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Previous studies have shown that the activities of NADH oxidase and cytoplasmic peroxidase on the yellow body membrane of rubber tree are enhanced, while the activities of SOD and CAT are weakened, and the concentration of antioxidants such as ascorbic acid and reduced

Method used

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  • Rubber plant translationally controlled tumor protein, encoding gene thereof and application of rubber plant translationally controlled tumor protein
  • Rubber plant translationally controlled tumor protein, encoding gene thereof and application of rubber plant translationally controlled tumor protein
  • Rubber plant translationally controlled tumor protein, encoding gene thereof and application of rubber plant translationally controlled tumor protein

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] Embodiment 1, the acquisition of HbTCTP protein and its coding gene

[0040] 1. Acquisition of cDNA of HbTCTP

[0041] By searching the established Hevea bark transcriptome deep sequencing database, a 803bp EST fragment of the translational control tumor protein gene was found. DNAstar7.0 software analysis showed that the EST fragment contained a complete open reading frame. Using Hevea brasiliensis Reyan 7-33-97 (planted in the test field of Chinese Academy of Tropical Agricultural Sciences and the National Rubber Tree Germplasm Resource Garden) as the experimental material, total RNA was extracted from its bark and reverse transcribed into cDNA. The cDNA was used as a template, and 5'-tgttgctgcttcttctctcct-3' and 5'-agaacaaaacaacaactatataaac-3' were used as primers for PCR amplification. The reaction mixture is as follows:

[0042] wxya 2 o

18.3μl

10×PCR buffer

2.5μl

dNTP Mixture (2.5mM)

2.5μl

Taq enzyme (5U / μl)

0.2μl...

Embodiment 2

[0052] Embodiment 2, functional verification of HbTCTP

[0053] 1. Prokaryotic expression and purification of HbTCTP

[0054] Primers OTLF: 5'-ggatccatgttggtctatcaggatttgc-3' and OTFR: 5'-gaattcttagcatttgacctccttcaaag-3' were designed according to the HbTCTP open reading frame, and the cDNA of the bark of Hevea Reyan 7-33-97 was used as a template for PCR amplification with high-fidelity enzymes , to obtain a PCR product of about 500bp.

[0055] Connect the above PCR product to the pMD18-T vector, transform E.coli DH5α competent cells, spread on the LB plate containing ampicillin (100mg / L), obtain a band of about 500bp by colony PCR, and identify the colony by PCR The monoclonal extracted plasmid was double-digested with EcoR I and BamHI to obtain a band of about 500bp, and the colony PCR and enzyme digestion identified positive clones to the company for sequencing to obtain the recombinant plasmid pMD18-HbTCTP. Use EcoR I and RanH I to cut out the target fragment on the rec...

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Abstract

The invention discloses a rubber plant translationally controlled tumor protein, an encoding gene thereof and an application of the rubber plant translationally controlled tumor protein. The translationally controlled tumor protein named HbTCTP (Hevea brasiliensis translationally controlled tumor protein) comes from Hevea brasiliensis which belongs to euphorbiaceae rubber category. The translationally controlled tumor protein is a protein (a) or a protein (b). The protein (a) is a protein which is composed of amino acid sequences showed in a second sequence of a sequence table. The protein (b) is a derivative protein, which is provided with antioxidant activity, of the second sequence by using one or more amino acid residues to substitute and/or delete and/or add the amino acid sequences. Experiments show that the rubber plant translationally controlled tumor protein can inhibit active oxygen cutting supercoiled plasmid DNA (deoxyribonucleic acid) generated by a metal catalytic oxidation system, so that the rubber plant translationally controlled tumor protein has antioxidant activity and can eliminate active oxygen in rubber plants.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a rubber tree translation control tumor protein and its coding gene and application. Background technique [0002] Natural rubber, together with steel, petroleum, and coal, is called the four major industrial raw materials. It is a basic industry and an important strategic material related to the national economy and people's livelihood. There are more than 2,000 kinds of rubber-producing plants in the world. Hevea brasiliensis Mull.Arg. has the advantages of high rubber production, good quality, long economic life, convenient rubber mining and low production cost, so it has become the only artificial The most important rubber-producing plants are cultivated, and the natural rubber produced by them accounts for more than 90% of the world's total natural rubber production. [0003] my country is a non-traditional rubber planting area, with limited rubber planting areas and relatively...

Claims

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Application Information

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IPC IPC(8): C07K14/415C12N15/29C12N15/63C12N5/10C12N1/15C12N1/19C12N1/21C12N15/11A01H5/00
Inventor 李德军邓治覃碧
Owner RUBBER RES INST CHINESE ACADEMY OF TROPICAL AGRI SCI
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