Cytosine methylation of plastid genome in higher plants.: Fact or artefact?

被引：39

作者：

Fojtová, M ^{[1
]}

Kovarík, A ^{[1
]}

Matyásek, R ^{[1
]}

机构：

[1] Acad Sci Czech Republ, Inst Biophys, CZ-61265 Brno, Czech Republic

来源：

PLANT SCIENCE | 2001年 / 160卷 / 04期

关键词：

DNA methylation; chloroplast; ribulose bisphosphate carboxylase/oxygenase; EcoRII restriction endonuclease;

D O I：

10.1016/S0168-9452(00)00411-8

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

DNA methylation of chloroplast genome has been studied in a large variety of angiosperm species using restriction enzyme analysis of three genomic loci (totally encompassing about 10% of chloroplast genome) and bisulfite genomic sequencing of tobacco ribulose bisphosphate carboxylase/oxygenase (large subunit) gene (rbcL). Except for CCWGG (W = A or T) sites that were partially refractory to the cleavage with methylation sensitive EcoRII in all loci. no cytosine methylation was found at the CCGG (dMspI/HpaII) and several other restriction sites tested. However, EcoRII was unable to completely digest an unmethylated CCWGG site in the cloned rbcL gene on plasmid. Further a bisulfite genomic sequencing performed on EcoRII-restricted DNA failed to show any 5-methylcytosine either within or outside inspected EcoRII sites along the 3 ' end of rbcL coding region. In conclusion our results do not support evidence for methylated cytosine residues in plant chloroplast genomes and we suggest that results obtained with EcoRII should be interpreted with great care especially when small differences in methylation levels are analysed. (C) 2001 Elsevier Science Ireland Ltd. All rights reserved.

引用

页码：585 / 593

页数：9

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