A Na<+>/H+ antiporter gene and applications thereof
A technology of antiporter protein and gene, applied in the field of genetic engineering, to achieve the effect of increasing accumulation and improving salt tolerance
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Embodiment 1
[0026] Example 1 Wheat plasma membrane Na + / H + antiporter gene TaSOS1-A1 the acquisition
[0027] 1. Wheat plasma membrane Na + / H + antiporter gene TaSOS1-A1 The separation specifically includes the following steps:
[0028] (1) Will wheat branch 26 ( Tritium aestivumL.) seeds were surface-sterilized with 10% Kao aqueous solution for 10 minutes, and washed 3 times with sterilized distilled water. Put the seeds into a Petri dish with two layers of sterilized filter paper, and after 2 days at room temperature (90% of the seeds germinated), put the Petri dish in a refrigerator at 4°C for 24 h to homogenize the germinated seeds. Then put the petri dish in a culture room (23°C, 16 h light / 8 h dark) to grow for 2 days, put the seedlings with consistent growth into a container containing 1 / 2 Hogland nutrient solution for hydroponics, and take the seedlings about 7 days later The leaves were washed and blotted dry, quick-frozen with liquid nitrogen, and stored in a -80°C...
Embodiment 2
[0035] Example 2 Wheat plasma membrane Na + / H + antiporter gene TaSOS1-A1 function of
[0036] 1. Construction of Dicotyledonous Binary Expression Vectors
[0037] Will TaSOS1-A1 The upstream primer of the gene plus the XbaⅠ restriction site 5´- GATCTAGAATGGAGACGGAGGAGGCCG-3´, the downstream primer is 5´- TCAGCTGCCTCGCGGTGG-3´), to TaSOS1-A1 The full-length T vector plasmid was used as a template for PCR amplification. The PCR products were recovered, connected to pGEM-T Vector, and then sequenced. The correctly sequenced plasmid TaSOS1-A1-T and the binary expression vector pCAMBIA1300-35S–GUS (referred to as p1300-GUS) were digested with XbaⅠ and SacⅠ, and the corresponding fragments were recovered, ligated and transformed; colony PCR was identified as the extraction of positive clones The plasmid was further identified by enzyme digestion, and the correctly identified plasmid p1300-TaSOS1-A1 was preserved for future use. The construction process of dicotyledonous ...
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