NtYpi1 gene capable of regulating and controlling potassium content of tobacco leaves and application of NtYpi1 gene
A technology of potassium content and tobacco leaves, applied in the field of genetic engineering, can solve the problems of cloning and research of tobacco protein phosphatase inhibitor which have not been seen before
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Embodiment 1
[0051] Example 1 Cloning of NtYpi1 Gene
[0052] 1.1 Experimental materials
[0053] 1.1.1 Tested strains
[0054] Escherichia coli DH5α is also preserved in our laboratory, and the applicant declares that it can be released to the public for verification experiments within 20 years from the date of application.
[0055] 1.1.2 Tobacco varieties tested
[0056] Tobacco: Yunyan 87, kept by the laboratory, the applicant declares that it can be released to the public for verification experiments within 20 years from the date of application.
[0057] 1.1.3 Test reagents
[0058] Phusion high-fidelity amplification enzyme reaction system, 5×Phusion HF reaction buffer, dNTP, Tris-HCl (PH8), EDTA, 20% SDS, β-mercaptoethanol, absolute ethanol, isopropanol, DNA extraction solution, 10× TAE and the above reagents were purchased from Sangon Biotechnology (Shanghai) Co., Ltd. (http: / / www.sangon.com). High-Fidelity DNA Polymerase was collected from Guangzhou Shengchuang Biotechnology ...
Embodiment 2
[0068] Example 2 Using CRISPR / cas9 to Knockout the NtYpi1 Gene to Reduce Potassium Content in Tobacco Leaves
[0069] 2.1 Experimental materials
[0070] 2.1.1 Tested strains
[0071]Escherichia coli DH5α. Agrobacterium (Agrobacterium) competent cells C58C1. The above biological materials are also preserved in this laboratory, and the applicant declares that they can be released to the public for verification experiments within 20 years from the date of application.
[0072] 2.1.2 Tobacco varieties tested
[0073] Tobacco: wild-type tobacco Yunyan 87, preserved in this laboratory, the applicant declares that it can be released to the public for verification experiments within 20 years from the date of application.
[0074] 2.1.3 Test reagents
[0075] pHSE401 vector, pK2GW7 vector, Trans-T1 competent cells, plasmid extraction kits, PCR product recovery kits, gel recovery kits, plant RNA extraction kits, and plant genome extraction kits were purchased from Beijing Quanshij...
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