Plant tillering related protein and coding gene thereof
A protein and gene technology, applied in the field of plant tiller-related proteins and their encoding genes
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Embodiment 1
[0040] Embodiment 1, the acquisition of OsMADS57 protein and its coding gene
[0041] Design primers according to the results of database analysis:
[0042] Overexpression vector construction using primer: 5' end primer: 5'-CG GGATCC ATGGGGAGGGGGAAGATAGT-3' (the underlined sequence is the BamHI site), 3' end primer: 5'-GG GGTACC TTAAGGCAGATGAAGTCCCAGT-3' (the underlined sequence is the KpnI site).
[0043] Antisense expression vector construction using primer: 5' end primer: 5'-CG GGATCC TTAAGGCAGATGAAGTCCCAGT-3' (the underlined sequence is the BamHI site), 3' end primer: 5'-GG GGTACC ATGGGGAGGGGGAAGATAGT-3' (the underlined sequence is the KpnI site).
[0044] Total RNA was extracted from the three-leaf stage seedlings of japonica rice Zhonghua No. 10, and the 726bp full-length cDNA of OsMADS57 was amplified by RT-PCR. The specific operation process is as follows:
[0045] 1) Extraction of total plant RNA: select 100 mg of Oryza sativa L.cvZhonghua 10 seedlings at th...
Embodiment 2
[0054] Embodiment 2, the application of OsMAOS57 protein and its coding gene
[0055] 1. Acquisition of overexpression vector pUN-MADS57 (sense) and antisense expression vector pUN-MADS57 (antisense)
[0056] 1. Acquisition of pUN1301 plasmid
[0057] Step 1: Cut about 0.2 g of corn seedlings and grind them in liquid nitrogen; then add 800 μL of freshly prepared extraction buffer (containing 0.1M Tris-HCl pH8.0, 50mM EDTA, 0.5M NaCl, 1% SDS and 1% β-mercaptoethanol), shake vigorously to suspend it completely; bathe in water at 65°C for 30 minutes, mix by inverting every 5 minutes; then add 250 μL of pre-cooled 5M potassium acetate aqueous solution, mix by inverting immediately, and bathe in ice for 5 minutes; Add an equal amount of phenol / chloroform, extract once, and centrifuge at 12,000rpm for 5 minutes; collect the supernatant, add 0.6 times the volume of isopropanol to precipitate DNA, and let it stand at room temperature for 40 minutes; centrifuge at 12,000rpm for 15 min...
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