Plant seed fatty acid related protein GhbZIP67 and coding gene and application thereof
A technology for plant seeds and fatty acids, applied in plant genetic improvement, botanical equipment and methods, applications, etc., can solve the relatively few studies on cottonseed oil content, the difficulty of cloning oil-related genes, and the slow progress of cottonseed oil-related gene location work, etc. question
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Embodiment 1
[0039] Embodiment 1, the acquisition of GhBZIP67 protein and its coding gene
[0040]The total RNA of different tissue and organ materials of cotton land-sea backcross inbred line 3008 was extracted and reverse transcribed into cDNA. After a large number of sequence analysis, expression analysis and functional verification, a DNA coding sequence was found from the cDNA, as shown in sequence 2 of the sequence listing, and its encoded protein was shown in sequence 1 of the sequence listing.
[0041] The protein shown in Sequence 1 of the Sequence Listing was named GhBZIP67 protein. The gene encoding GhBZIP67 protein was named as GhBZIP67 gene.
Embodiment 2
[0042] Example 2, Functional verification of GhBZIP67 protein and its coding gene
[0043] 1. Obtaining of transgenic plants
[0044] 1. Construction of the recombinant expression vector: The fragment between the BamHI and SacI restriction sites of the pBI121 vector was replaced with the DNA molecule shown in the sequence 2 of the sequence listing from the 1-1176 position of the 5' end to obtain the recombinant expression vector pBI121: : GhBZIP67 (sequencing verification).
[0045] 2. The recombinant expression vector pBI121::GhBZIP67 obtained in step 1 was introduced into Agrobacterium GV3101 to obtain the recombinant strain GV3101::GhBZIP67.
[0046] 3. Transfer the recombinant strain GV3101::GhBZIP67 obtained in step 2 into Arabidopsis Col-0 by the flower dipping method. The specific transformation steps are as follows:
[0047] (1) Inoculate the recombinant bacteria GV3101::GhBZIP67 obtained in step 2 into YEB liquid medium containing 100mg / L rifampicin, 25mg / L Genta an...
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