Salt resistance gene msvp2 of energy crop hag and its plant expression vector and construction method
A technology for plant expression vector and salt tolerance gene, applied in the field of molecular biology, can solve the problems of limited salt tolerance, low efficiency, affecting cultivation and application, etc., and achieve the effect of improving salt tolerance and variety improvement
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[0034] Example 1 Construction and Transformation of Plant Expression Vector pEarleyGate103-MsVP2
[0035] 1. Cloning of MsVP2:
[0036] Grass (Miscanthus sacchariflora) was selected as the material, and healthy cuttings were selected, and the seedlings (20 days) that survived the cuttings were continuously stressed with salt (200mM NaCl) for 3 days, and 0.05g of young leaves were taken, and were extracted with reference to the Trizol RNA extraction kit (TaKaRa ) method described in the instructions, extracting total RNA from leaves, taking 1 μg of total RNA to reverse transcribe into cDNA according to the M-MLV Reverse Transcription Kit (TaKaRa), digesting the cDNA product with RNase, referring to the Arabidopsis VP2 sequence information (NCBI accession number: AT1G16780 ), analyzed and designed primers to amplify MsVP2 by Oligo6.0 software;
[0037] Upstream primer MsVP2-F: 5′-ATGATGGAAGAAGACGTGGA-3′
[0038] Downstream primer MsVP2-R: 5′-CAAGAATATTGGTGCCATGACC-3′
[0039]...
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