Bidirectional promoter bi-visual fluorescent protein report gene plant expression vector
A technology for expressing vectors and foreign proteins, applied in the direction of using vectors to introduce foreign genetic material, microorganisms, microorganisms, etc., can solve problems such as promoter silencing, vector rearrangement or deletion, and interaction, and achieve the effect of easy screening
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[0018] Embodiment 1, construction general type plant bidirectional promoter expression vector
[0019] Construction of general-purpose plant bidirectional promoter expression vector
[0020] On the basis of the pBI121 vector, the p131 plasmid obtained by replacing the gus gene with the gfp gene, using the p131 plasmid as a template, and using D1 and D2 as primers, PCR amplifies a sequence from Pmini to the terminator:
[0021] D1: 5'-AAGCTTGGTACCCCCTTCGCAAGACCCTTC-3',
[0022] D2: 5'-GAAGCTTGTCACTGGATTTTGGTTTTAGG-3'.
[0023] PCR amplification conditions were: pre-denaturation at 94°C for 5mins; 94°C for 30s, 67°C for 30s, 72°C for 1min, 25 cycles; 72°C for 10mins; storage at 4°C.
[0024] The PCR product was recovered, connected with pMD18 T-Vector, and the cloning vector pT-MGN was constructed, and blue-white screening was carried out on LB medium. Pick a few white spots and culture them, extract the plasmids, digest with HindIII for preliminary verification, and send the...
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