Metal transporter protein and encoding gene and application thereof

A metal and utility technology, applied in the direction of genetic engineering, plant genetic improvement, application, etc., can solve the problem of poor understanding of the mechanism of iron absorption and transportation

Active Publication Date: 2013-10-30
CAS CENT FOR EXCELLENCE IN MOLECULAR PLANT SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The biofortification method using molecular breeding to increase the iron content in edible parts such as crop grains has good and broad prospects for development, but the key theoretical issue involved in biofortificat

Method used

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  • Metal transporter protein and encoding gene and application thereof
  • Metal transporter protein and encoding gene and application thereof
  • Metal transporter protein and encoding gene and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0127] Embodiment 1, OsVIT1 and OsVIT2 gene sequence and structure

[0128] The inventor cloned two genes from rice: OsVIT1 (LOC_Os04g0463400) and OsVIT2 (LOC_Os09g0396900), wherein the gene sequence and its structure are as follows:

[0129] OsVIT1 (SEQ ID NO: 1, obtained from Start coding, underlined is the coding region): AAATAAACCACCTCTTCAAACGTGCGTCGCCTGACCGCTTCCATCCATCCACCAGTCTCGCTTCTTCGTGTCTACGGTCTACCTCGGCGAGTCGGCGGCGGCG GCGGCGGCGACGGACGGCGGGGGGCTGC CGCTGCTGGCGGACAAGGCGGCGAGCCACAGCCACCACCACCACCCGGAGCGGCACTTCACGTCGG GGGAGGTGGTCCGCGACGTCATCATGGGCGTCTCCGACGGCCTCACCGTGCCCTTCGCCCTCGCCGC GGGGCTCTCCGGCGCCAGCGCGCCATCCTCGCTCGTGCTCACCGCCGGCCTCGCCGAGGTCGCCGCC GGCGCCATCTCCATGGGCCTCGGAGGGTAAACATTAAACACTCATCATCTCTCTGTGTGTTCCTTGTC CATGCATTGCACCCCTGAATCCCTGATTAATCTAGCGAAGTTAGCAATATGTCAGTTTCGTCG tatagatttgctcttcttttcttttttttttaaagaggggttccgagtttttcattaccaaaacaggaaacccagcggcagggtgcagtagtatgtcagtgaatacaatagaaagctttcggggacccacaggccacagcccataacatgatactagtaatttattaatcacctgaaatcg...

Embodiment 2

[0141] Example 2. Transcription levels of OsVIT1 and OsVIT2 genes in different tissues of rice and their responses to iron content in the environment

[0142] In this example, real-time fluorescent quantitative RT-PCR was used to analyze the transcription and expression of rice OsVIT1 and OsVIT2 genes under different iron treatment or iron deficiency treatment conditions.

[0143] Real-time quantitative fluorescent RT-PCR primer sequence:

[0144] OsVIT1(+) RT-PCR: TCCTCAAGCTTGGCTCGACTTCAT (SEQ ID NO: 7);

[0145] OsVIT1(-) RT-PCR: ATGGCGTTCTGTGCTGTGGAGATA (SEQ ID NO: 8);

[0146] OsVIT2(+) RT-PCR: GGCTGCAGGCATCCAAGTAAATGT (SEQ ID NO: 9);

[0147] OsVIT2(-) RT-PCR: CACTACAAGCACGCAGCAAACGTA (SEQ ID NO: 10);

[0148] OsACT1(+) RT-PCR: TGGAAGCTGCGGGTATCCAT (SEQ ID NO: 11);

[0149] OsACT1(-) RT-PCR: TACTCAGCCTTGGCAATCCACA (SEQ ID NO: 12).

[0150] Primers were synthesized by Shanghai Sangon Bioengineering Technology Service Co., Ltd. OsACT1 (OsActin1), a constitutively expr...

Embodiment 3

[0159] Example 3, OsVIT for yeast Fe, the impact of Zn sensitive mutants

[0160] The present inventors speculate that the proteins encoded by OsVIT1 and OsVIT2 have the function of transporting Fe(II) from the cytoplasm to the vacuole. To test this inference, the present inventors investigated the functions of OsVIT1 and OsVIT2 by yeast complementation experiments.

[0161] The yeast Fe-sensitive mutant used in this example refers to the document Localization of Iron in Arabidopsis Seed Requires the Vacuolar Membrane Transporter VIT1.SCIENCE.314.1295 (2006). Yeast Zn-sensitive mutants refer to the literature Induction of the ZRC1 Metal Tolerance Gene in Zinc-limited Yeast Confers Resistance to Zinc Shock. THE JOURNAL OF BIOLOGICAL CHEMISTRY. 278.15065 (2003). Yeast cadmium (Cd) sensitive mutant (Δycf1) refers to literature A Yeast Metal Resistance Protein Similar to Human Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) and Multidrug Resistance-associated Protein. ...

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Abstract

The invention relates to a metal transporter protein and an encoding gene and an application thereof. The invention discloses a new gene; and polypeptide encoded by the new gene has a metal transporter function. The gene can be greatly applied to the improvement of plant varieties and also can be used for increasing the content of iron and/or zinc in edible parts of a plant. The invention provides a valuable gene resource for developing a new crop variety with higher nutritive value.

Description

technical field [0001] The invention belongs to the field of biotechnology; more specifically, the invention relates to a metal transporter, its encoding gene and its use. Background technique [0002] Iron is an essential trace element for the human body. Iron deficiency can lead to various diseases in the human body. The most common and most harmful one is iron deficiency anemia. Iron deficiency anemia is recognized in the world as one of the four major nutritional deficiency diseases that are more serious and widespread. According to statistics, about 3 billion people in the world are iron deficient, and the incidence of anemia caused by iron deficiency is as high as 30% of the world's total population (http: / / www.who.iht / nutrition / topics / ida / ), of which People in developing countries are most severely deficient in iron. One of the main reasons is that people in developing countries generally eat cereals as their staple food, but the iron content in cereals is low, and ...

Claims

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Application Information

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IPC IPC(8): C07K14/415C12N15/29C12N15/63C12N1/15C12N1/19C12N1/21C12N5/10C12P21/02A01H5/00
Inventor 龚继明张雨
Owner CAS CENT FOR EXCELLENCE IN MOLECULAR PLANT SCI
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