a na + /h + Retrotransporter gene and its application

A technology of reverse transport proteins and genes, applied in the field of genetic engineering, to achieve the effect of improving salt tolerance and increasing accumulation

Inactive Publication Date: 2020-07-28
SHIJIAZHUANG ACADEMY OF AGRI & FORESTRY SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Mutations in this gene in Arabidopsis make the plant very sensitive to salt and cause Na + Efflux and Na + Defects in long-distance transport from roots to stems

Method used

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  • a na <sup>+</sup> /h <sup>+</sup> Retrotransporter gene and its application
  • a na <sup>+</sup> /h <sup>+</sup> Retrotransporter gene and its application
  • a na <sup>+</sup> /h <sup>+</sup> Retrotransporter gene and its application

Examples

Experimental program
Comparison scheme
Effect test

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 the 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 -...

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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Abstract

The invention discloses a kind of Na on the wheat plasma membrane + / H + Antiporter genes and their applications. The Na + / H + The full length of the antiporter gene is 3460bp, and after expression, the gene is located on the plasma membrane of the cell and plays a role in Na + / H + exchange function, the Na + out of the cell, and thus the present invention also discloses the use of the Na + / H + The antiporter gene constructs a plant genetic transformation vector and a method for improving the salt tolerance of the plant.

Description

technical field [0001] The invention relates to the field of genetic engineering, in particular to a + extracellular Na + / H + Antiporter genes and their applications. Background technique [0002] First cloned from Arabidopsis in 2000 SOS1 genes and are thought to be located on the plasma membrane at the Na + / H + Antiporter protein has 12 stringed membrane domains at its N-terminus and a long hydrophilic tail at its C-terminus. The results of subcellular localization showed that the protein is localized on the cell membrane, and it is the only protein responsible for Na in plants identified on the plasma membrane. + efflux protein. Na + / H + antiporter converts H + Na + transported outside the cell. Mutations in this gene in Arabidopsis make the plant very sensitive to salt and cause Na + Efflux and Na + Defects in long-distance transport from roots to stems and leaves. Normally, SOS1 will make SOS1 dormant through the interaction between the C-terminal self...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/29C12N15/81C12N15/83C12N15/84C12N15/11
CPCC07K14/415C12N15/81C12N15/8203C12N15/8205C12N15/8273C12N2800/102C12N2800/60
Inventor 高振贤张爱民史占良阳文龙刘冬成郭进考
Owner SHIJIAZHUANG ACADEMY OF AGRI & FORESTRY SCI
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