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Paddy rice zinc finger protein gene and its coded protein

A zinc finger protein and genetic engineering technology, applied in the field of genetic engineering, can solve the problems that there is no functional description of rice C2H2 type zinc finger protein gene, and the relationship between WZF1 stress response has not been found yet.

Inactive Publication Date: 2007-06-20
NANJING AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Although the monocot C2H2-type zinc finger protein WZF1 was first reported in wheat, the relationship between WZF1 and stress response has not been found
Rice is an important food crop and a model plant for monocots. So far, there is no functional description of the rice C2H2 zinc finger protein gene

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Molecular Cloning of Rice Zinc Finger Protein Gene OsZFP18

[0025] The rice variety "Jiucaiqing" (japonica rice local variety in the Taihu Lake Basin, a strong salt-tolerant variety) was selected. After the rice seedlings grew to the 3-leaf stage, they were treated with 140mM NaCl, and the leaves were immediately taken out and frozen in liquid nitrogen after 12 hours. . Take part of the leaves, grind them with a mortar, add them into a 1.5mLEP tube filled with lysate, oscillate fully, and then transfer them into a glass homogenizer. After homogenization, transfer to 1.5mLEP tube, and extract total RNA (TRIzolReagents, Invitrogen, USA). The quality of total RNA was identified by formaldehyde denaturing gel electrophoresis, and then the RNA content was determined on a spectrophotometer. The GenBank rice genome database was searched with the EST sequence S121 from the rice salt-induced cDNA library, and the full-length 883bp sequence of the rice C2H2-type zinc finger pr...

Embodiment 2

[0028] Sequence information and characteristic analysis of OsZFP18

[0029] The full length of OsZFP18 of the present invention is 883bp, and its open reading frame is located at positions 53-568. DNAssist software analysis showed that OsZFP18 encoded a total of 171 amino acids, which contained 2 typical Cys2 / His2 zinc finger structures, and the Cys-Xaa-Cys and His-Xaaa-His regions were composed of 12 mainly acidic or hydrophobic amino acids The coordination structure plus the hydrophobic core formed by Phe and Leu determine the basic structure and shape of the zinc finger. Both zinc finger structures contain the conserved structure of QALGGH. By Blast program comparison, it was found that the amino acid similarities between the encoded protein of OsZFP18 and soybean zinc finger protein SCOF-1, wheat WZF1, and Arabidopsis STZ were 33%, 36%, and 30%, respectively. Its pI=5.19 and MW=17.9 KDa were estimated using Compute pI / MW software. Searching the existing rice genome (inc...

Embodiment 3

[0031] Expression study of OsZFP18

[0032] Use the primers at both ends of OsZFP18 designed in Example 1 to carry out semi-quantitative RT-PCR analysis of the expression of the aboveground and underground parts of rice seedlings, and use the expression of rice actin gene Racl (McElroy et al, 1990) as an internal reference. The results show that rice seedlings OsZFP18 The expression of 140mM NaCl was significantly enhanced at 6h, and the expression reached the highest at 24h. Gray-scale scanning analysis showed that its expression was about 10 times that of the control.

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Abstract

The invention discloses a first nucleotide sequence and protein sequence coded by it of paddy C2H2 type zinc finger protein gene OsZFP18 that belongs to gene engineering field. The OsZFP18 anticipates in the development of the fringe and the salt menace response. The expression of mRNA analysis indicates it is induced by high salt. The invention also discloses the method to improve the salt endurance of the plant by using OsZFP18.

Description

technical field [0001] The invention relates to a genetic engineering application of a rice zinc finger protein gene OsZFP18, which belongs to the field of genetic engineering, and in particular relates to a zinc finger transcription factor responding to plant salt stress. technical background [0002] C2H2-type zinc finger proteins widely exist in various plants and participate in plant growth and development and environmental stress response (Huang Ji et al., 2004). Nearly 30 C2H2-type zinc finger protein genes have been cloned in Petunia and Arabidopsis. The research results show that they are mainly involved in the development of plants in the reproductive growth stage, or in abiotic processes such as high salinity, low temperature, and drought. stress response. The study of soybean zinc finger protein SCOF-1 showed that overexpression of SCOF-1 in Arabidopsis and tobacco significantly enhanced the expression of cold-inducible protein genes, thereby improving the cold t...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/29C07K14/415
Inventor 王建飞黄骥张红生王州飞郭书巧
Owner NANJING AGRICULTURAL UNIVERSITY
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