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

Inactive Publication Date: 2016-10-26
SHIJIAZHUANG ACADEMY OF AGRI & FORESTRY SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Mutations in this gene in Arabidopsis make the plant very sensitive to salt an...

Method used

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  • A Na&lt;+&gt;/H+ antiporter gene and applications thereof
  • A Na&lt;+&gt;/H+ antiporter gene and applications thereof
  • A Na&lt;+&gt;/H+ antiporter gene and applications thereof

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 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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PUM

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Abstract

A Na<+>/H+ antiporter gene on a wheat plasma membrane and applications thereof are disclosed. The full length of the antiporter gene is 3460 bp. The gene after expression is positioned on a cell plasma membrane, and exerts a Na<+>/H+ exchange function to discharge Na<+> out of cells. A method of constructing a plant genetic transformation vector by utilizing the antiporter gene and improving plant salt tolerance is also disclosed.

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 sel...

Claims

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Application Information

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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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