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通过两个含有同源序列的抑制基因座实现植物中转基因的差异失活和甲基化。

Differential inactivation and methylation of a transgene in plants by two suppressor loci containing homologous sequences.

作者信息

Matzke M A, Matzke A J

机构信息

Institute of Molecular Biology, Austrian Academy of Sciences, Salzburg.

出版信息

Plant Mol Biol. 1991 May;16(5):821-30. doi: 10.1007/BF00015074.

Abstract

In a previous study on doubly transformed tobacco plants, we observed the unexpected inactivation in trans of T-DNA-I (encoding KanrNOS) following the introduction into the same genome of an unlinked copy of T-DNA-II (encoding HygrOCS). This inactivation, which probably resulted from interactions between homologous regions on each T-DNA, was correlated with methylation in the nospro, which controlled the expression of both the nptII and nos genes. In this paper, we show that the inactivation and methylation of the nos(pro)nptII gene in the presence of a suppressor T-DNA-II locus can be either complete (epistasis) or partial (cellular mosaicism). In plants showing partial suppression, the strength of the Kanr phenotype, which apparently reflected the proportion of cells expressing the nptII gene, was inversely correlated with the degree of methylation of the nos(pro). The extent of nos(pro) methylation decreased progressively in successive generations as suppressor T-DNA-II loci were crossed out. The strength of the Kanr phenotype was improved and nos(pro) methylation was less extensive in first generation Kanr progeny obtained from outcrossing with untransformed tobacco than from self-fertilization.

摘要

在之前一项关于双转化烟草植株的研究中,我们观察到,在将编码潮霉素抗性基因(HygrOCS)的T-DNA-II的非连锁拷贝导入同一基因组后,编码卡那霉素抗性基因(KanrNOS)的T-DNA-I出现了意外的反式失活。这种失活可能是由于每个T-DNA上同源区域之间的相互作用导致的,并且与nos启动子(nospro)的甲基化相关,该启动子控制着nptII和nos基因的表达。在本文中,我们表明,在存在抑制性T-DNA-II位点的情况下,nos(pro)nptII基因的失活和甲基化可以是完全的(上位性)或部分的(细胞镶嵌性)。在表现出部分抑制的植株中,卡那霉素抗性(Kanr)表型的强度显然反映了表达nptII基因的细胞比例,它与nos(pro)的甲基化程度呈负相关。随着抑制性T-DNA-II位点在连续世代中被剔除,nos(pro)的甲基化程度逐渐降低。与自交获得的第一代卡那霉素抗性(Kanr)后代相比,与未转化烟草杂交获得的第一代卡那霉素抗性(Kanr)后代中,卡那霉素抗性(Kanr)表型的强度有所提高,并且nos(pro)的甲基化程度也较低。

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