Homology-dependent gene silencing in Paramecium
Homology-dependent gene silencing in Paramecium
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DOI：
10.1091/mbc.9.4.931
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发表时间：
1998-04-01
影响因子：
3.3
通讯作者：
Madeddu, L
中科院分区：
文献类型：
--
作者：
Ruiz, F;Vayssie, L;Madeddu, L
将仅含基因编码区的质粒以高拷贝数显微注射到草履虫体细胞大核中，导致相应内源基因的表达显著降低。沉默的效果，这是稳定地保持在整个营养生长，已被观察到的所有草履虫基因检查到目前为止：单拷贝基因（ND7），以及多基因家族的成员（中心蛋白基因和包囊基质蛋白基因），其中所有密切相关的旁系同源基因似乎受到影响。这一现象可能与转基因植物的转录后基因沉默和脉孢菌的抑制有关，并允许有效地产生特定的突变表型，从而为研究草履虫的基因功能提供了一个潜在的强大工具。对于编码共同组装以建立复杂亚细胞结构的蛋白质的两个多基因家族，本文提供的分析提供了第一个实验证据，表明这些基因家族的成员在功能上并不冗余。
Microinjection at high copy number of plasmids containing only the coding region of a gene into the Paramecium somatic macronucleus led to a marked reduction in the expression of the corresponding endogenous gene(s). The silencing effect, which is stably maintained throughout vegetative growth, has been observed for all Paramecium genes examined so far: a single-copy gene (ND7), as well as members of multigene families (centrin genes and trichocyst matrix protein genes) in which all closely related paralogous genes appeared to be affected. This phenomenon may be related to posttranscriptional gene silencing in transgenic plants and quelling in Neurospora and allows the efficient creation of specific mutant phenotypes thus providing a potentially powerful tool to study gene function in Paramecium. For the two multigene families that encode proteins that coassemble to build up complex subcellular structures the analysis presented herein provides the first experimental evidence that the members of these gene families are not functionally redundant.