Plant nitrogen utilization efficiency related protein TaNF-YA1 and coding gene and application thereof
A technology for encoding genes and proteins, which can be used in plant gene improvement, application, plant peptides, etc., and can solve the problems of difficult functional gene research and large genome in wheat.
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[0048] Embodiment 1, cultivating transgenic plants and detection thereof
[0049] 1. Breeding transgenic plants
[0050] 1. Acquisition of gene (TaNF-YA1)
[0051] The total RNA of wheat Xiaoyan 54 (Henan Zhongke Seed Industry Co., Ltd.) was extracted, and the whole genome cDNA of wheat was obtained by reverse transcription. As a template, the upstream primer with XbaI restriction site: 5'-CT TCTAGA ATGCTTCTCCCCTCTTCT-3' and downstream primer with BamHI restriction site: 5'-TT GGATCC TTACGTATACCGGTCGCTCGA-3' was used as a primer for PCR amplification. The PCR system was 40 μl in total, including 4 μl of template cDNA, 1 μl of KOD-plus-DNA polymerase (purchased from TOYOBO Company), 10 × PCR buffer for KOD-plus-4 μl, dNTPs ( 2mM each) MgSO 4 (25mM) upstream and downstream primers The reaction system was supplemented with double distilled water to The PCR reaction program was: 98°C for 2min, (98°C for 30sec, 58°C for 30sec, 68°C for 45sec)×35cycles. After the react...
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