Establishment of gene-trap and enhancer-trap systems in the moss Physcomitrella patens
Establishment of gene-trap and enhancer-trap systems in the moss Physcomitrella patens
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DOI：
10.1046/j.1365-313x.2001.01121.x
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发表时间：
2001-10-01
期刊：
影响因子：
7.2
通讯作者：
Hasebe, M
中科院分区：
文献类型：
--
作者：
Hiwatashi, Y;Nishiyama, T;Hasebe, M
小立碗藓由于其简单的体型和易于基因敲除和等位基因替换，常被用作植物生理学和发育生物学研究的模式系统。基因捕获和增强子捕获系统是用于克隆在特定组织或细胞中起作用的基因和增强子的有用技术。此外，这些系统便于获得对某些发育过程特异的分子标记。使用具有剪接受体或最小启动子的uidA报告基因构建基因陷阱和增强子陷阱系统的元件。通过同源重组的方法获得了高转化率，从5637个和3726个转基因株系中分别获得了235个基因诱捕株系和1073个增强子诱捕株系。这些诱捕线在苔藓配子体中的表达模式各不相同。通过5'端和3'端RACE技术，从假根特异表达的基因诱捕系YH 209中筛选到候选基因。该基因被命名为PpGLU，与植物酸性α-葡萄糖苷酶基因形成一个进化枝.因此，这些基因陷阱和增强子陷阱系统应该证明是有用的，以确定组织和细胞特异性基因立碗藓。
Because of its simple body plan and ease of gene knockout and allele replacement, the moss Physcomitrella patens is often used as a model system for studies in plant physiology and developmental biology. Gene-trap and enhancer-trap systems are useful techniques for cloning genes and enhancers that function in specific tissues or cells. Additionally, these systems are convenient for obtaining molecular markers specific for certain developmental processes. Elements for gene-trap and enhancer-trap systems were constructed using the uidA reporter gene with either a splice acceptor or a minimal promoter. Through a high rate of transformation conferred by a method utilizing homologous recombination, 235 gene-trap and 1073 enhancer-trap lines were obtained from 5637 and 3726 transgenic lines, respectively. The expression patterns of these trap lines in the moss gametophyte varied. The candidate gene trapped in a gene-trap line YH209, which shows rhizoid-specific expression, was obtained by 5' and 3' RACE. This gene was named PpGLU, and forms a clade with plant acidic alpha -glucosidase genes. Thus, these gene-trap and enhancer-trap systems should prove useful to identify tissue- and cell-specific genes in Physcomitrella.