Plant senescence-related protein GhWRKY91 as well as coding gene and application thereof
A plant senescence and plant technology, applied to the plant senescence-related protein GhWRKY91 and its coding gene and application field, can solve the problems of short-season cotton premature senescence, etc., and achieve the effect of great application value
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[0061] 1. Construction of recombinant expression vector
[0062] 1. Extract the mature leaves of the upland cotton Zhongmiansuo No. 74 plant at the full flowering stage, and extract the total RNA.
[0063] 2. Reverse transcribe the total RNA obtained in step 1 to obtain cDNA.
[0064] 3. Using the cDNA obtained in step 2 as a template, perform PCR amplification with a primer pair composed of F1 and R1, and recover the PCR amplification product.
[0065] F1: 5'-CTAG TCTAGA ATGGACAGCGGTGGTAG-3';
[0066] R1: 5'-TCC CCCGGG TCAGCAGATCCCAACCTG-3'.
[0067] 4. Digest the PCR amplified product obtained in step 3 with restriction endonucleases XbaI and SmaI, and recover the digested product.
[0068] 5. Digest the pBI121 plasmid with restriction endonucleases XbaI and SmaI to recover the vector backbone.
[0069] 6. Ligate the digested product obtained in step 4 with the vector backbone obtained in step 5 to obtain recombinant plasmid pBI121-GhWRKY91. According to the sequenc...
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