Wheat heat stress associated protein TaGCN5 as well as coding gene and application thereof
A gene and protein technology is applied to the wheat heat stress related protein TaGCN5 and its encoding gene and application field, which can solve the problems of grain yield decrease, quality deterioration, influence on grain filling, protein and starch synthesis, etc., to increase the ability of high temperature stress tolerance Effect
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Embodiment 1
[0034] Example 1. Discovery of wheat heat stress-related protein TaGCN5 and its encoding gene
[0035] On the basis of the NCBI database, the genes and fragments with related functions from other plants were compared, and according to the results of comparison and sequence analysis, the relevant fragments of wheat were spliced, extended and functionally identified, and finally a The completed open reading frame encodes the protein shown in Sequence 1 of the Sequence Listing, and the protein is named TaGCN5 protein. The gene encoding TaGCN5 protein is named as TaGCN5 gene, as shown in sequence 2 of the sequence listing.
Embodiment 2
[0036] Embodiment 2, the acquisition and identification of transgenic plants
[0037] 1. Construction of recombinant plasmids
[0038] 1. Synthesize the double-stranded DNA molecule shown in sequence 2 of the sequence listing.
[0039] 2. Using the double-stranded DNA molecule synthesized in step 1 as a template, perform PCR amplification with a primer pair composed of F1 and R1 to obtain a PCR amplification product.
[0040] F1: 5'-CACCCGG GGTACC ATGGACGGCCTCGCGGCGCC-3';
[0041]R1: 5'-CTGGGTC GAGCTC CTAGTTCTTCGTTGACGCAG-3'.
[0042] 3. The PCR amplified product of step 2 was double-digested with restriction endonucleases KpnI and SacI, and the digested product was recovered.
[0043] 4. Digest the pCAMBIA-1300 vector with restriction endonucleases KpnI and SacI to recover the vector backbone.
[0044] 5. Ligate the digested product of step 3 with the vector backbone of step 4 to obtain a recombinant plasmid. According to the sequencing results, the structure of the ...
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