HbsHsp1 protein coming from hevea brasiliensis, and coding gene and application thereof
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A coding and gene technology, applied in the field of HbsHsp1 protein derived from rubber tree and its coding gene and application, can solve the problem of rubber tree yield reduction and other problems
Inactive Publication Date: 2014-03-26
RUBBER RES INST CHINESE ACADEMY OF TROPICAL AGRI SCI
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my country is a non-traditional rubber planting area, and rubber trees are often affected by abiotic adversity conditions such as rubber tapping, low temperature, drought, and wind damage, resulting in a decrease in the output of rubber trees
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Embodiment 1
[0036] Embodiment 1, the acquisition of HbsHsp1 protein and its coding gene
[0037] Established a deep sequencing database of the transcriptome of the rubber tree bark, and found a 632bp EST fragment from Hevea brasiliensis Hot Research 7-33-97. The EST fragment, as shown in sequence 2 of the sequence listing, contains a complete open reading frame and encodes the The protein shown in sequence 1.
[0038] The protein shown in Sequence 1 of the Sequence Listing is named as HbsHsp1 protein. The gene encoding the HbsHsp1 protein is named as the HbsHsp1 gene, and its open reading frame is shown in the 28th-522nd nucleotides from the 5' end of the sequence 2 of the sequence listing.
[0039]The bark, latex, leaves, male flowers, female flowers and anthers of Hevea brasiliensis Reyan 7-33-97 were taken respectively, and the total RNA was extracted and reverse-transcribed into cDNA, and the relative expression of HbsHsp1 gene was detected by using a primer pair consisting of F1 and...
Embodiment 2
[0043] Example 2, Functional Verification of HbsHsp1 Protein
[0044] 1. Construction of recombinant plasmids
[0045] 1. Extract the total RNA from the bark of Hevea brasiliensis Reyan 7-33-97 and reverse transcribe to obtain cDNA.
[0046] 2. Using the cDNA obtained in step 1 as a template, perform PCR amplification with a primer pair composed of F2 and R2 to obtain a PCR amplification product.
[0049] 3. Digest the PCR amplified product obtained in step 2 with restriction endonucleases BamH I and EcoR I, and recover the digested product.
[0050] 4. Digest the pET28a(+) vector with restriction endonucleases BamH I and EcoR I to recover a vector backbone of about 5400 bp.
[0051] 5. Ligate the digested product obtained in step 3 with the vector backbone obtained in step 4 to obtain the recombinant plasmid pET28a-HbsHsp1. According to the sequencing results,...
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Abstract
The invention discloses HbsHsp1 protein coming from hevea brasiliensis, and coding gene and application thereof. The provided protein is the following (a) or (b) or (c): (a) protein comprising the amino acid sequence shown in the sequence 1 of a sequence table; (b) protein derived from the protein of the sequence 1 by substituting and/or deleting and/or adding one or multiple amino acid residues related to the anti-oxidation capability of the plate from the amino acid sequence of the sequence 1; and (c) protein derived from the protein of the sequence 1 by substituting and/or deleting and/or adding one or multiple amino acid residues related to the anti-oxidation capability of microbes from the amino acid sequence of the sequence 1. The provided protein participates in the oxidative stress resistant response of plants or microbes. The provided protein and the coding gene thereof play an important role for improving the oxidative stress resistant capability of plants or microbes and improving hevea brasiliensis yield or reducing hevea brasiliensis dead bark.
Description
technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a HbsHsp1 protein derived from a rubber tree, its coding gene and its 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 characteristics of high rubber production, good quality, long economic life, convenient rubber mining and low production cost, so it has become the only artificial The cultivated rubber-producing plants account for more than 90% of the world's total natural rubber production. [0003] The Brazilian rubber tree is native to the Amazon River Basin in South America and is a typical tropical tall tree. my country is a non-traditional ...
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