Salt Du's algal nitrate transfer protein and its code sequence

A Dunaliella nitrate and nitrate transport technology, applied in the field of biology, can solve problems such as laborious, time-consuming and lack of pertinence

Inactive Publication Date: 2005-06-08
四川川大光耀生物工程有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, traditional methods of plant improvemen...

Method used

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0081] Cloning of Nitrate Transporter Gene of Dunaliella salina

[0082] 1. Collection of Dunaliella salina

[0083] Dunaliella salina (Dunaliella salina) was purchased from the algae bank of Wuhan Institute of Hydrobiology.

[0084] 2. Poly A + RNA isolation (Poly A + RNA isolation)

[0085] Total RNA of Dunaliella salina was extracted with Trizol reagent (Gibco, NY, USA). The quality of total RNA was identified by formaldehyde denaturing gel electrophoresis. Dunaliella salina mRNA was extracted using the kit instructions provided by Oligotex mRNA Kits (Qiagen).

[0086] 3. Construction of Dunaliella salina cDNA library (Cloning of Full-length cDNA)

[0087] The cDNA phage library of Dunaliella salina was constructed using the kit instructions provided by the Smart cDNA Library Construction Kit (ClonTech) with λTriplEX2 as the vector.

[0088] 4. Cloning of Full-length cDNA

[0089] By comparing the nitrate transporters of a large number of different species (includin...

Embodiment 2

[0150] Structure and function of a nitrate transporter in Dunaliella salina

[0151] The full-length sequence of Dunaliella salina nitrate transporter gene and its encoded protein were analyzed by BLAST program in Non-redundant GenBank+EMBL+DDBJ+PDB and Non-redundant GenBank cdstranslations+PDB+SwissProt+PIR databases for nucleotide and protein Homology search revealed that it has 45% identity and 61% similarity with Chlamydomonas reinhardtii nitrate transporter (GenPept Accession No. AJ223296.2).

[0152]The amino acid of the Dunaliella salina nitrate transporter was searched for the structural domain in the Conserved Domain Database (http: / / www.ncbi.nlm.nih.gov / Structure / cdd / cdd.shtml), and it was found that the amino acid sequence was the most The conserved domain of the long-reading frame protein is similar to the conserved domain of the nitrate transporter in the protein sequence 49-401aa. It is generally believed that the membrane protein of this family contains twelve t...

Embodiment 3

[0179] Large-scale sequencing of Dunaliella salina cDNA library and Construction of Expression Microarray

[0180] The clones in the obtained Dunaliella salina cDNA library were sequenced, and the detected target sequences were used to construct an expression profile chip. The clones obtained by sequencing were dissolved in 3×SSC solution, and the Cartesian 7500 spotting instrument of Cartesian Company and the silanized glass slide of TeleChem Company were used to operate according to the instructions provided by the company. After sample application, the slides were hydrated, dried, and UV cross-linked, then treated with SDS and water for 10 minutes, and then dried for later use.

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Abstract

A novel salt resistance associated protein-Du's saline alga nitrate transporter, the polynucleotide for coding it, the process for preparing said protein by recombination, the application of said polynucleotide, and the method for improving the salt resistance of plant by use of said transporter are disclosed.

Description

technical field [0001] The invention belongs to the field of biology. Specifically, the invention relates to a new polynucleotide encoding a salt-tolerance-related protein-nitrate transporter of Dunaliella salina, and a polypeptide encoded by the polynucleotide. The present invention also relates to the use and preparation of such polynucleotides and polypeptides. Specifically, the polypeptide of the present invention is a new salt-tolerance-related protein. Background technique [0002] Currently, the world's population is increasing while the arable land is decreasing. However, there are still many land resources on the earth that cannot be effectively used due to salinization. In order to effectively utilize these salinized land resources, people have been looking for plant varieties that are both adaptable to growth and have high economic value. However, no very suitable plant species have hitherto been available. [0003] Another method for developing saline-alkali-...

Claims

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Application Information

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IPC IPC(8): A01H1/00C07H21/00C07K14/405C12N15/63C12N15/79C12P21/00
Inventor 曹毅蒋彦唐克轩杨滔曾昌耀
Owner 四川川大光耀生物工程有限公司
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