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Protein arginine methyltransferase of Arabidopsis thalianum and its coding gene and application

A technology of arginine methylation and protein, applied in the direction of transferase, application, genetic engineering, etc., can solve the problem that there is no report on histone arginine methylation, achieve high practical application value, and improve tolerance , the effect of broad application prospects

Inactive Publication Date: 2007-01-03
INST OF GENETICS & DEVELOPMENTAL BIOLOGY CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although eight PRMTs have been identified in mammalian cells, they play an important role in regulating chromatin remodeling, gene transcription, signal transduction and RNA splicing (Bedford MT, Richard S. Arginine methylation: an emerging regulator of protein function.Mol Cell.2005,18(3):263-72.), but there is no report on the role of PRMT and its catalyzed histone arginine methylation in plant development and stress response

Method used

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  • Protein arginine methyltransferase of Arabidopsis thalianum and its coding gene and application
  • Protein arginine methyltransferase of Arabidopsis thalianum and its coding gene and application
  • Protein arginine methyltransferase of Arabidopsis thalianum and its coding gene and application

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Experimental program
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Embodiment 1

[0035] Embodiment 1, the cloning of the gene AtPRMT5 of Arabidopsis protein arginine methyltransferase

[0036] The obtaining of the full-length cDNA sequence of the gene of Arabidopsis protein arginine methyltransferase comprises the following steps:

[0037] For the human PRMT5 gene sequence (GenBank No.: NM_006109) published in GenBank, a homologous sequence (GenBank No.: NM_119262) was obtained from the Arabidopsis genome by comparison of homologous sequences, and according to the 5' and Design a pair of primers for the 3' end sequence, the primer sequence is as follows:

[0038] P1 (upstream primer); 5'-aagagctccatgccgctcggagagagaggagg-3'

[0039] P2 (downstream primer): 5'-aactcgagaaggccaacccagtacgaacgg-3'

[0040] The total RNA of Arabidopsis leaves was extracted (Promega, SV total RNA isolation system), and the total RNA of Arabidopsis leaves was used as a template to synthesize cDNA with AMV reverse transcriptase (TaKaRa) (according to Plant RT-PCR Kit2.01 (TaKaRa ...

Embodiment 2

[0041] Example 2. Detection of the regulatory effect of AtPRMT5 on the flowering and development of Arabidopsis thaliana

[0042] 1. Screening of AtPRMT5 inactivation homozygous mutants

[0043] Live homozygous mutants of Arabidopsis AtPRMT5 were screened by the following method: the seeds (Salk 065814 and Salk 095085) of two different T-DNA insertion mutant lines at the AtPRMT5 site were obtained from ABRC (Arabidopsis Biological Resource Center) (Alonso, J.M., Stepanova, A.N., Leisse, T.J., Kim, C.J., Chen, H., Shinn, P., Stevenson, D.K., Zimmerman, J., Barajas, P., Cheuk, R., et al. 2003. Genome- wide insertion mutagenesis of Arabidopsisthaliana.Science 301:653-657.), two kinds of AtPRMT5 inactivation homozygous mutants with different T-DNA insertion sites were obtained by PCR identification, named AtPRMT5-1 and AtPRMT5-2 respectively.

[0044] 2. Comparison of the growth of AtPRMT5 inactivation homozygous mutants and wild-type plants

[0045] Harvest the seeds of the AtP...

Embodiment 3

[0051] Example 3. Detection of the influence of AtPRMT5 on histone amino acid modification

[0052] The antibody Anti-H4-R3-sdiMe, which can specifically recognize the symmetrical double-methylation modification of the third arginine position of histone H4 (purchased from Jingmei Bioengineering Company, ABCAM Company) was used to detect the homozygous mutant of AtPRMT5 inactivation and wild-type plants (control, WT) were detected by Western blot, the results were as follows figure 2 As shown, the expression levels of histone methylation in AtPRMT5 inactivation homozygous mutants and wild-type plants are significantly different, indicating that the inactivation of AtPRMT5 leads to symmetrical methylation of the third arginine of histone H4 The level is reduced, affecting the covalent modification of other sites of histone amino acids, changing the codon of histone, thereby affecting the transcription and expression of key genes (such as FLC) in the plant flowering signal transd...

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Abstract

The present invention discloses one protein arginine methyl transferae of Arabidopsis thalianum and its coding gene and application in regulating and controlling growth and resistance of plant. The protein arginine methyl transferae has one of the following amino acid residue sequence: 1) SEQ ID No. 1 in the sequence list; and 2. the amino acid residue sequence of SEQ ID No. 1 through substitution, deletion or addition of 1-30 amino acid residues and coding protein regulating and controlling growth and resistance of plant. Experiment shows that the gene of the present invention can raise the growth speed and high salt stress tolerance of plant obviously, and the present invention provides one high quality gene for improvement of plant variety and possesses high practical application and wide application foreground.

Description

technical field [0001] The present invention relates to plant protein arginine methyltransferase and its encoding gene and its application, in particular to protein arginine methyltransferase and its encoding gene from Arabidopsis thaliana and its role in regulating plant growth and development and stress resistance sexual applications. Background technique [0002] Physiological processes such as flowering development and environmental stress response in plants are regulated by multiple genes. Many botanists at home and abroad have preliminarily revealed the gene regulatory network of plant flowering development and environmental stress response. Studies have shown that the flowering development time of Arabidopsis is mainly regulated by four main pathways: autonomy, vernalization, gibberellin and photoperiod (He Y, Michaels SD, Amasino RM Regulation of flowering time by histone acetylation in Arabidopsis. Science, 2003 302: 1751-1754). The research on the gene regulator...

Claims

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Application Information

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IPC IPC(8): C12N9/10C12N15/54C12N15/63C12N5/10C12N15/82
Inventor 鲍时来种康王昕张娅徐云远马其斌
Owner INST OF GENETICS & DEVELOPMENTAL BIOLOGY CHINESE ACAD OF SCI
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