Corn CCCH-type zinc finger protein, and encoding gene ZmC3H54 and application thereof

A technology of zinc finger protein and coding gene, which is applied in application, genetic engineering, plant gene improvement, etc., to achieve the effects of improving tolerance, improving stress resistance, and improving the ability to resist abiotic adversity stress

Inactive Publication Date: 2015-08-12
ANHUI AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] At present, there is a lack of a maize CCCH-type zinc finger protein ZmC3H54 and its coding gene ZmC3H54 with strong ability to resist abiotic stress

Method used

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  • Corn CCCH-type zinc finger protein, and encoding gene ZmC3H54 and application thereof
  • Corn CCCH-type zinc finger protein, and encoding gene ZmC3H54 and application thereof
  • Corn CCCH-type zinc finger protein, and encoding gene ZmC3H54 and application thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0059] Acquisition of ZmC3H54 Gene in Maize

[0060] After the maize seedlings grew to the three-leaf stage, total RNA was extracted and reverse transcribed into cDNA. The MAIZEGENOME and NCBI databases were searched to obtain the deduced coding sequence of ZmC3H54. Primer Premier 5.0 software was used to design specific primers, which were synthesized by Sangon Biotechnology. The primer sequences are as follows:

[0061] ZmC3H54-F 5′-CG GGATCC ATGTACAACTTCAAAGTGAAGC-3'SEQ ID NO.5

[0062] BamHI

[0063] ZmC3H54-R 5′-GC TCTAGA TCAGGCCACCATCTGCTC-3'SEQ ID NO.6

[0064] XbaI

[0065] Using the above-obtained maize total cDNA as a template, a coding region containing a complete open reading frame was obtained by PCR with a length of 1119 bp, which was recovered, connected to the pEASY-T1 vector, and sequenced. The resulting sequence is the maize ZmC3H54 gene.

[0066] Transformation of plant expression vector pCAMBIA1301

[0067] The...

Embodiment 2

[0088] Expression pattern analysis and tissue pattern analysis of maize ZmC3H54 gene under PEG, ABA, NaCl conditions

[0089] like image 3 Shown is the analysis of the induced expression pattern of the ZmC3H54 gene of the present invention under ABA, drought, high-salt stress treatment and the tissue expression pattern of the ZmC3H54 gene; A: 10% PEG solution treatment; B: 100uM ABA solution treatment; C: 200mmol / L NaCl solution treatment;

[0090]1. Sampling under stress treatment: the plant material is the corn B73 inbred line. When the corn seedlings grow to the three-leaf stage, the plants with normal irrigation are used as the control, and 10% PEG solution, 100uM ABA solution and 200mmol / L NaCl solution are respectively carried out. For stress treatment, samples were taken after treatment for different time periods, respectively 0h, 1h, 3h, 6h, 12h, and 24h; the leaves of the same part of the maize seedlings in the experimental group and the control group were cut with ...

Embodiment 3

[0133] Sequence homology and homology analysis of maize ZmC3H54 gene

[0134] like figure 1 Shown, is the amino acid sequence homology comparison map of ZmC3H54 of the present invention and other species CCCH family zinc finger protein conserved regions; The zinc finger protein family has the most recent homology relationship. The transcription factor was compared with the protein sequences of other known members of the CCCH type zinc finger protein family, and the zinc finger structure type was analyzed as CCCH type. According to the structural characteristics of the DNA binding domain, the structure of the ZmC3H54 protein was CX 8 CX 5 CX 3 H.

[0135] like figure 2 shown, is the phylogenetic tree after alignment of the amino acid sequences of ZmC3H54 of the present invention and the CCCH zinc finger protein family with reported functions in Arabidopsis and other plants; in order to further analyze the relationship between ZmC3H54 and other CCCH zinc finger proteins wi...

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Abstract

The invention discloses a corn CCCH-type zinc finger protein, and an encoding gene ZmC3H54 and an application thereof. An amino acid sequence of the corn CCCH-type zinc finger protein is as shown in SEQ ID NO.1 in a sequence table. The nucleotide acid sequence of the encoding gene ZmC3H54 of the corn CCCH-type zinc finger protein disclosed by the invention is as shown in SEQ ID NO.2 in the sequence table. The invention further discloses a recombinant carrier, a recombinant bacterium, a transgenic cell line or an expression cassette containing the encoding gene ZmC3H54. The ZmC3H54 gene disclosed by the invention participates into stress response, so that the resistance to abiotic adversity stress of a plant can be improved; a transgenic plant has relatively high sensitivity on ABA; the tolerance of transgenic rice on drought is improved; the family gene is separated and cloned from corn; and the developing function of the family gene in the aspect of improving the stress resistance of a target plant is identified.

Description

technical field [0001] The invention belongs to the field of crop genetics and breeding, and in particular relates to a corn CCCH type zinc finger protein and its encoding gene ZmC3H54 and application. Background technique [0002] Corn is an important food crop and an important source of feed, and it is also the food crop with the highest total yield in the world. Corn is the most suitable variety for industrial raw materials among the three major food crops, and occupies an important position in the national economy. However, abiotic stresses such as drought and high salinity seriously affect the yield of maize. Therefore, discovering and creating new germplasms and breeding new varieties of maize with strong stress resistance, high yield and high quality have become the key to the sustainable development of maize production. With the rapid development of molecular biology and plant genetic engineering breeding technology, genetic engineering breeding technology to improv...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/415C12N15/29C12N15/63C12N1/15C12N1/19C12N1/21C12N15/82A01H5/00C12Q1/68
CPCC07K14/415C12N15/8273C12N15/8293C12Q1/6851C12Q2545/113
Inventor 朱苏文徐倩倩赵阳彭元成彭晓剑
Owner ANHUI AGRICULTURAL UNIVERSITY
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