Methylation of coding region alone inhibits gene expression in plant protoplasts
Methylation of coding region alone inhibits gene expression in plant protoplasts
复制标题
DOI：
10.1073/pnas.93.16.8334
复制
发表时间：
1996-08-06
影响因子：
11.1
通讯作者：
Rothnie, H
中科院分区：
文献类型：
--
作者：
Hohn, T;Corsten, S;Rothnie, H
花椰菜花叶病毒35 S启动子的衍生物缺乏CG和CNG甲基化的目标仅仅构建和用于指导瞬时转化的原生质体中的报告基因构建体的转录。这样的无甲基化靶标（MTF）启动子，虽然弱于35 S启动子，但尽管as-1元件突变，仍保留显著的活性。检测了甲基化对MTF和35 S启动子驱动构建体中基因表达的影响。即使当启动子区是免费的甲基化目标，报告基因的表达显着降低时，在CG二核苷酸中的胞嘧啶残基在体外甲基化转化前。嵌合甲基化实验，其中仅质粒的特定部分被甲基化，揭示了单独的编码区的甲基化对报告基因表达具有负面影响。靠近编码区5'端的甲基化更具抑制性，与转录延伸的抑制一致。
Derivatives of the cauliflower mosaic virus 35S promoter lacking CG and CNG methylation targets mere constructed and used to direct transcription of reporter gene constructs in transiently transformed protoplasts. Such methylation-target-free (MTF) promoters, although weaker than the 35S promoter, retain significant activity despite mutation of the as-1 element. The effect of methylation on gene expression in MTF- and 35S-promoter driven constructs was examined. Even when the promoter region was free of methylation targets, reporter gene expression was markedly reduced when cytosine residues in CG dinucleotides were methylated in vitro prior to transformation. Mosaic methylation experiments, in which only specific parts of the plasmids were methylated, revealed that methylation of the coding region alone has a negative effect on reporter gene expression, Methylation nearer the 5' end of the coding region was more inhibitory, consistent with inhibition of transcription elongation.