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Complementary deoxyribonucleic acid (cDNA) sequence related to iron deficiency of plant and encoded protein and application thereof

A plant, iron deficiency technology, applied in the field of plant genetic engineering, can solve problems affecting absorption, anemia, economic loss, etc.

Inactive Publication Date: 2012-06-13
CAPITAL NORMAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Iron deficiency in plants not only affects the growth and development of plants, causing huge economic losses, but also affects the absorption of edible iron in animals and humans, which leads to the occurrence of anemia (Guerinot ML, et al.1994)

Method used

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  • Complementary deoxyribonucleic acid (cDNA) sequence related to iron deficiency of plant and encoded protein and application thereof
  • Complementary deoxyribonucleic acid (cDNA) sequence related to iron deficiency of plant and encoded protein and application thereof
  • Complementary deoxyribonucleic acid (cDNA) sequence related to iron deficiency of plant and encoded protein and application thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0035]Example 1 Cloning of the full sequence of maize ZmFDR

[0036] 1. RNA extraction

[0037] The iron-sensitive maize variety (Zea mays L cv Yedan 12) was surface sterilized with 0.5%-1% NaOCl, and then germinated at 27°C for 36h. When the root length was about 1 cm, corn 11-24d was cultured (-Fe culture) at 25° C. with a liquid without ferrite (Hoagland broth). When the symptoms of iron deficiency appeared, about 2 cm of the root tip was cut every day and stored at -80°C for later use. Mixed and random samples were taken when building the bank. Simultaneously control corn is in normal iron content (concentration is 1x10 -4 mol / L) culture solution (+Fe culture).

[0038] 2. Construction of DNA library

[0039] Press ZAP Express TM cDNA Synthesis Kit and ZAP Express TM Cdna Gigapack GoldII Clong Kit instructions for cDNA library construction. First-strand synthesis is anchored with an adapter-primer containing an Xho I site:

[0040] 5'GAGAGAGAGAGAGAGAGAGAACTAGTCTC...

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Abstract

The invention discloses a complementary deoxyribonucleic acid (cDNA) sequence related to iron deficiency of a plant and an encoded protein and application thereof. A full-length cDNA of a ZmFDR gene is cloned from an iron-deficient induced corn root and the structural characteristic of the full-length cDNA and the functional complementation of the full-length cDNA in yeast are analyzed. The analysis of the structural characteristic indicates that the full length of the ZmFDR gene is 3.3 kb, the gene comprises two open reading frames which are 1,068 bp and 648 bp respectively and the latter isnamed as ZmFDR4 (SEQ ID NO:1). The encoded protein (SEQ ID NO:2) has four transmembrane domains and has a signal peptide which belongs to a secretory protein and has a similar structure domain to a Flip protein in an III type secretory system. Extensive homology comparison indicates that no homology sequence is found in a higher plant. A yeast functional complementation experiment proves that theexpression of the cDNA sequence in the yeast can transfer iron ions effectively.

Description

technical field [0001] The present invention relates to a cDNA sequence related to plant mineral nutrition and its encoded protein, especially to a cDNA sequence related to corn iron deficiency and its encoded protein. The present invention also relates to a recombinant expression vector containing the cDNA sequence and the cDNA The use of the sequence in improving plant resistance to iron deficiency stress belongs to the field of plant genetic engineering. Background technique [0002] Iron is the earliest trace element discovered by people, and it has been discovered for more than two thousand years. Iron has an irreplaceable function in the life activities of prokaryotes and eukaryotes existing on the earth. The study of iron in plants can be traced back to 1843, a century and a half ago, when Gris found that the chlorosis of grape leaves growing on calcareous soil was related to iron deficiency. After research by botanists Sachs and Molisch, it was identified as an ess...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/29C12N15/63C12N15/82C12N1/19C07K14/415A01H5/00
Inventor 印莉萍韩建辉闫佳洁马小娟关丽英宋秀芳张艳萍霍春雁白彦霞刘祥林
Owner CAPITAL NORMAL UNIVERSITY
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