Histone demethylase gene OsJMJ706 of rice, encoding protein thereof and applications

A demethylase and histone technology, applied in the field of plant genetic engineering, can solve problems such as little understanding of the regulation of flower organ development in monocotyledonous plants

Inactive Publication Date: 2009-02-11
HUAZHONG AGRI UNIV
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  • Abstract
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Problems solved by technology

[0006] At present, the molecular mechanism of genetic regulation of flower organ development in dicotyledonous plants has been basically elucidated, but little is known about the regulation of floral organ development in monocotyledonous plants

Method used

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  • Histone demethylase gene OsJMJ706 of rice, encoding protein thereof and applications
  • Histone demethylase gene OsJMJ706 of rice, encoding protein thereof and applications
  • Histone demethylase gene OsJMJ706 of rice, encoding protein thereof and applications

Examples

Experimental program
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Effect test

Embodiment 1

[0029] Example 1: Co-segregation detection and expression analysis of mutant plants

[0030] 1. Rice material

[0031]The original wild-type material of rice (Oryza sativa ssp.) used in this example is a rice variety "Zhonghua 11" japonica rice variety (ZH11) publicly promoted and applied in China, and it is also a rice T-DNA insertion mutant database (http: / / rmd.ncpgr.cn / ) is a receptor material used. In order to find the T-DNA insertion mutant of OsJMJ706, the applicant used the OsJMJ706 gene sequence (see the gene sequence of the present invention cloned as shown in SEQ ID NO: 1) in the rice T-DNA insertion mutant database (http: / / rmd.ncpgr .cn / ) website analysis tool (http: / / rmd.ncpgr.cn / blast.cgi?nickname=) for BLAST analysis (Zhang J et al., a rice mutant database for functional analysis of the rice genome.Nucl.Acids Res., 2006, 34: D745-748), found two allelic mutants, the original numbers were 03Z11F083 and 05Z11D089, for convenience, the applicant renamed jmj6-1 (0...

Embodiment 2

[0055] Example 2: Analysis of histone modification in mutants

[0056] Since the OsJMJ706 gene contains the jmjC and jmjN domains unique to the Jumonji family, and its human homolog gene JMJD2 family gene has been reported to have demethylase activity at histone H3K9 and H3K36 sites, in order to detect whether the OsJMJ706 gene has demethylase activity For the activity of the base enzyme, the applicant used the western blot method (referring to "Molecular Cloning Experiment Guide", written by J. Sambrook and D.W. Russell, translated by Huang Peitang, Science Press, 2002) to the wild-type ZH11 and mutants. Histone modifications in jmj6-2 were detected in vivo. Specific steps are as follows:

[0057] 1. Histone extraction method

[0058] Take 4-5 leaves of wild-type ZH11 and mutant jmj6-2 about 40 days after sowing, grind them with liquid nitrogen and put them into 3 times the volume of histone extraction buffer (10mM Tris-HCl, pH7.5; 2mM EDTA; 0.25M HCl; 5mM 2-mercaptoethano...

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Abstract

The invention belongs to the technical field of plant gene engineering, and more particularly relates to a paddy rice histone demethyl methylase gene, coded protein and the application thereof. The invention provides a new paddy rice histone demethyl methylase gene OsJMJ706, due to inactivation, heteroplasia of rice flowerthe, heteroplasia or deficit of internal and external glumes, as well as lessening of seeds can be caused on the gene, and by being detected by the method of western blot, the gene is proved to have enzyme activity for removing histone H3K9me3 and H3K9me2 locus methylate. The invention provides nucleic acid sequence and protein sequence of the gene as well as the application thereof.

Description

technical field [0001] The invention relates to the technical field of plant genetic engineering, in particular to a rice histone demethylase gene, its encoded protein and its application. The present invention provides a new rice histone demethylase gene OsJMJ706. The inactivation of this gene can lead to abnormal development of rice flowers, abnormal development or loss of inner and outer glumes, and smaller seeds, which can be detected by western blot method. It has enzymatic activity to demethylate histone H3K9me3 and H3K9me2 sites. The present invention provides the nucleic acid sequence of the gene, its protein sequence and its application. Background technique [0002] Histone modification-related enzymes play an important role in the growth and development of animals and plants, such as common heterosis, nucleolar dominance, vernalization, Paramutation, transgene silencing, transposon activation, genomic imprinting, DNA methylation , RNA editing, and X chromosome i...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/52C12N9/00A01H1/00A01H5/00
Inventor 周道绣孙前文
Owner HUAZHONG AGRI UNIV
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