HbsHsp1 protein coming from hevea brasiliensis, and coding gene and application thereof
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
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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.
[0047] F2: 5'-gc ggatcc atgtcgatcaatcccttcttc-3';
[0048] R2: 5'-gc gaattc tcaaccagaaatcttaatatagac-3'.
[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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