Efficient gene targeting in the moss Physcomitrella patens
Efficient gene targeting in the moss Physcomitrella patens
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DOI：
10.1046/j.1365-313x.1997.11061195.x
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发表时间：
1997-06-01
期刊：
影响因子：
7.2
通讯作者：
Zryd, JP
中科院分区：
文献类型：
--
作者：
Schaefer, DG;Zryd, JP
利用单倍体配子体在其生命周期中占主导地位的事实，苔藓小立碗藓被用作研究植物发育的遗传模型系统。进行了旨在靶向三个单拷贝基因组基因座的转化实验，以确定该植物中基因靶向的效率。靶向载体的平均转化率提高了 10 倍，并且提供了通过同源重组将外源 DNA 整合到每个靶向位点的分子证据。这些实验确定的基因打靶效率超过90%，处于酵母中观察到的范围内，比之前报道的植物基因打靶高出几个数量级。因此，基因敲除和等位基因替换方法可直接用于研究小立碗藓植物的发育。此外，迄今为止，仅在原生动物、酵母和丝状真菌等低等真核生物中观察到有效的基因靶向，并且如此处所示，来自植物界的第一个例子是单倍体苔藓。这表明有效的基因靶向与真核生物的单倍体之间可能存在相关性。
The moss Physcomitrella patens is used as a genetic model system to study plant development, taking advantage of the fact that the haploid gametophyte dominates in its life cycle. Transformation experiments designed to target three single-copy genomic loci were performed to determine the efficiency of gene targeting in this plant. Mean transformation rates were 10-fold higher with the targeting vectors and molecular evidence for the integration of exogenous DNA into each targeted locus by homologous recombination is provided. The efficiency of gene targeting determined in these experiments is above 90%, which is in the range of that observed in yeast and several orders of magnitude higher than previous reports of gene targeting in plants. Thus, gene knock-out and allele replacement approaches are directly accessible to study plant development in the moss Physcomitrella patens. Moreover, efficient gene targeting has so far only been observed in lower eukaryotes such as protozoa, yeasts and filamentous fungi, and, as shown here the first example from the plant kingdom is a haplobiontic moss. This suggests a possible correlation between efficient gene targeting and haplophase in eukaryotes.