Rice secretary type thioredoxin gene and application thereof
A thioredoxin, secreted technology, applied in the field of genetic engineering, can solve problems such as plant aging, crop yield reduction, oxidative damage, etc., and achieve the effects of reducing hydrogen peroxide content, delaying rice aging, and increasing crop yield
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[0019] 1. Acquisition of genes
[0020] This thioredoxin is a protein that the applicant found during two-dimensional electrophoresis, and the expression of rice root apoplasts is up-regulated under salt stress conditions. The Accession No. of the protein is gi|3915131, and its gene sequence number is AK121423. In order to obtain the gene, the applicant extracted the RNA of wild-type rice (Oryza sativa), and used primers 5'-CCATGGCCGCCGAGGAGGGA-3' and 5'-CCATGGCGGCAGAAGCAGATGCAGCAGTG-3' for RT-PCR amplification. The reaction system was as follows: ultrapure water 31ul, 10×Pyrobest Buffer II 5ul, dNTP Mixture 4ul, upstream and downstream primers 4ul, Pyrobest DNA Polymerase 0.2ul, cDNA 2ul. The PCR program is: 5 minutes at 94°C, 30 seconds at 94°C, 30 seconds at 61°C, 50 seconds at 72°C, 27 cycles from the second to fourth steps, and 10 minutes at 72°C. The PCR product was subjected to electrophoresis, recovered from the gel, connected to the T carrier, and the full-length nuc...
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