Protein PwRBP1 related to stress tolerance of plants, and encoding gene and application thereof
A protein and plant technology, applied in the direction of plant gene improvement, application, plant peptides, etc., can solve the problems of few reports on RBP gene function.
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Embodiment 1
[0078] Embodiment 1, the acquisition of PwRBP1 and its coding gene
[0079] 1. Acquisition of PwRBP1 and its coding gene
[0080] In this paper, using the cDNA library of Cinnamon cDNA constructed by the Gateway method completed by Yingwei Jieji (Shanghai) Co., Ltd. as a template, 5'-CTGATGAATATCGACTTTGGAAAGTC-3' and 5'-ATGATGCAACCCACAGCAGG-3' were used for PCR amplification to obtain positive cDNA sequence, and sequenced.
[0081] The sequencing results show that the cDNA nucleotide sequence is as shown in SEQ ID NO.1 in the sequence listing, consisting of 1293 nucleotides, the gene is named PwRBP1, the protein encoded by PwRBP1 is named PwRBP1, and its amino acid sequence is the sequence listing Among them, SEQ ID NO.2 consists of 431 amino acids.
[0082] The above cDNA can be obtained by artificial synthesis.
[0083] 2. Fluorescent quantitative PCR analysis of tissue expression of PwRBP1 gene
[0084] RNA from pollen, roots, stems, needles, and seeds of T. chinensis w...
Embodiment 2
[0086] Embodiment 2, the application of PwRBP1 gene in improving plant stress tolerance
[0087] 1. Construction of transgenic PwRBP1 Arabidopsis
[0088] 1. Acquisition of PwRBP1 gene
[0089] Prepare the following amplification primer pairs:
[0090]Primer 1: 5'-ATGATGCAACCCACAGCAGG-3';
[0091] Primer 2: 5'-CTGATGAATATCGACTTTGGAAAGTC-3'.
[0092] Using the cDNA of P. chinensis as a template, the above primers 1 and 2 were used for PCR amplification to obtain a PCR product with a size of 1293 bp, which was sequenced after being connected with the pEASY-T1 vector.
[0093] Sequencing results show that the nucleotide sequence of the PCR product is SEQ ID NO.1 in the sequence listing, which can encode the protein PwRBP1 shown in SEQ ID NO.2 in the sequence listing.
[0094] 2. Acquisition of recombinant expression vector
[0095] Prepare the following primer pairs:
[0096] Primer 1: 5'-CGCGGATCCATGATGCAACCCACAGCAGG-3';
[0097] Primer 2: 5'-TCCCCCGGGCTGATGAATATCGACTTTGG...
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