Arsenic-resistance related protein PvArrp1, and coding gene and application thereof
A resistance-related, genetic technology, applied in application, genetic engineering, plant genetic improvement, etc., to achieve the effect of enhancing resistance, great value, and reducing the accumulation of arsenic
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Embodiment 1
[0047] Embodiment 1, the invention of PvArrp1 protein and its coding gene
[0048] Centipede grass (Pteris vittata L.), belongs to the genus Pteris vittata of the fern family Pteridaceae.
[0049] The inventor discovered a protein related to arsenic resistance from Centipede grass, and named it PvArrp1 protein. The amino acid sequence of PvArrp1 protein is shown in sequence 1 of the sequence listing. The gene encoding PvArrp1 protein is named PvArrpl gene, and its open reading frame is shown in sequence 2 of the sequence listing.
Embodiment 2
[0050] Embodiment 2, the acquisition and identification of transgenic plants
[0051] 1. Construction of recombinant expression vector
[0052] 1. Extract the total RNA of the centipede grass blade.
[0053] 2. Reverse transcribe the total RNA extracted in step 1 into cDNA.
[0054] 3. Using the cDNA obtained in step 2 as a template, perform PCR amplification with a primer pair composed of PvArrp1S and PvArrp1A to obtain a PCR amplification product.
[0055] PvArrp1S: 5'-ATGGAGAACTCAAGCGCGGAGCGGA-3';
[0056] PvArrp1A: 5'-CTAAACAGAAGGCCCCTTCCTCTGA-3'.
[0057] PCR amplification program: 98°C for 10s, 55°C for 30s, 72°C for 70s, 35 cycles; 72°C for 10min; store at 10°C.
[0058] 4. Using the PCR amplification product in step 3 as a template, a primer pair composed of PvArrp1S1 / PvArrp1A1 is used to obtain a PCR amplification product.
[0059] PvArrp1S1: 5'- ggatcc ATGGAGAACTCAAGCGCGGAGCGGA-3';
[0060] PvArrp1A1: 5'- ggtacc CTAAACAGAAGGCCCCTTCCTCTGA-3'.
[0061] T...
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