Promoter capable of driving expression of gene in tissue of flower
A promoter and expression vector technology, applied in the field of promoters, can solve the problems of normal elongation of filaments, hindered stamen development, male sterility, etc., achieve broad application space and market prospects, and promote development and growth.
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Embodiment 1
[0025] Example 1, the acquisition of MYB21pro sequence
[0026] Using Arabidopsis thaliana (Columbia-0) leaf DNA as a template, using a primer pair composed of F1 and R1, PCR amplification was performed under the action of high-fidelity PFU enzyme to obtain PCR amplification products.
[0027] F1:5'-agaAAGCTTataattactccacctt-3':
[0028] R1: 5'-tttTCTAGAtttagagagaggaatat-3'.
[0029] The PCR amplification product was sequenced, as shown in sequence 1 of the sequence listing. The DNA shown in Sequence 1 of the Sequence Listing was named MYB21 promoter (MYB21pro).
Embodiment 2
[0030] Embodiment 2, the acquisition and identification of transgenic plants
[0031] 1. Construction of recombinant expression vector (pBI121-MYB21pro-GUS)
[0032] 1. The PCR amplification product obtained in Example 1 was double-digested with restriction endonucleases HindIII and XbaI to obtain the digested product.
[0033] 2. The pBI121 vector (GenBank: AF485783.1) was double digested with restriction endonucleases HindIII and XbaI, and the vector backbone of about 14 kb was recovered.
[0034] 3. Ligate the digested product of step 1 and the vector backbone of step 2 under the action of T4 DNA ligase to obtain the recombinant plasmid pBI121-MYB21pro-GUS. The schematic diagram of the structure of the recombinant plasmid pBI121-MYB21pro-GUS is shown in figure 1 . According to the sequencing results, the structure of the recombinant plasmid pBI121-MYB21pro-GUS is described as follows: the recombinant plasmid obtained by inserting the DNA shown in sequence 1 in the sequen...
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