Protein AtLPT4 related to deficient-phosphorus stress tolerance of plants, and coding gene and application thereof
A technology for low-phosphorus tolerance and plants, which is applied to the protein AtLPT4 related to plant resistance to low-phosphorus stress and its encoding gene and application field, which can solve problems such as unclear functions
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Embodiment 1
[0047] Embodiment 1, acquisition and detection analysis of transgenic AtLPT4 plants
[0048] 1. Obtaining transgenic AtLPT4 Arabidopsis
[0049] 1. Acquisition of cDNA
[0050] Total RNA was extracted from Arabidopsis thaliana (Columbia type (Col-0 ecotype), (purchased from the American Arabidopsis Biological Resource Center, ABRC)) seedlings for 10 days, and reverse-transcribed into cDNA. The cDNA was Template, utilize respectively to be provided with a pair of primers of XbaI and KpnI restriction site to carry out PCR amplification, obtain the cDNA full-length of the gene AtLPT4 of plant resistance to low phosphorus stress related protein.Primer sequence is as follows:
[0051] Primer1: 5′-GC tctaga ATGGAGGATAGGAGGTGTGA-3' (XbaI);
[0052] Primer2: 5′-gg ggtacc TCATTCCTTCAAGCAAAAGGGA-3' (KpnI).
[0053] The high-fidelity Pyrobest DNA polymerase from Takara Company was used for amplification. The amplification system was 50 μL, containing 5.0 μL of 10×Pyrobest PCR buff...
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