RNAi (RNA interfere) plant expression vector of soybean allergy protein gene Gly m Bd 28K
A plant expression vector and protein technology, applied in DNA/RNA fragments, plant products, recombinant DNA technology, etc., can solve the problems of limited utilization space, time-consuming, and difficulty in changing soybean quality traits at the same time, and achieve plant variety improvement, The effect of eliminating the harm of allergic people
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[0055] 1. Transformation of pBI121 vector
[0056] 1) Transformation of the nos sequence of pBI121
[0057] ①Design primers P1 and P2, and use pBI121 plasmid as a template to amplify the nos terminator fragment containing the Asc I site,
[0058] Upstream primer P1: 5`-AGGCGCGCCGATCGTTCAAACATTTGGCA-3`
[0059] Downstream primer P2: 5`-GGAATTCGATCTAGTAACATAGATGACACC-3`
[0060] Amplification system: template pBI121 plasmid 1.0 μL (-1 ), dNTPmix 4.0μL (2.5mmol·L -1 ), PrimeSTAR TM HS DNA Polymerase 0.2 μL, ddH 2 O 37.8 μL;
[0061] Amplification program: pre-denaturation at 94°C for 4min, denaturation at 94°C for 30s, annealing at 51.9°C for 30s, extension at 72°C for 30s, extension at 72°C for 10min, 30 cycles;
[0062] The PCR product was connected into the cloning vector pMD19-T simple vector and verified by sequencing;
[0063] ② Design primer P3, and downstream primer P2, use the nos terminator fragment containing Asc I site as a template to amplify the nos fragment...
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