Andreeva A V, Kutuzov M A, Evans D E, Hawes C R
Research School of Biological & Molecular Sciences, Oxford Brookes University, Oxford, 0X3 OBP, U.K.
Cell Biol Int. 1998;22(2):145-60. doi: 10.1006/cbir.1998.0235.
Numerous proteins have been identified in yeast and mammalian cells which are involved in trafficking between the endoplasmic reticulum and the Golgi apparatus. A great number of partial cDNA sequences now available from the two major plant model species, Arabidopsis thaliana and Oryza sativa, makes it possible to identify putative plant homologues of known genes/proteins from non-plant species. The authors used this approach to screen the database of Expressed Sequence Tags (dbEST) in order to detect plant homologues of proteins involved in membrane transport between ER and Golgi. Availability of these partial sequences will facilitate the screening of cDNA and genomic libraries otherwise performed using heterologous probes derived from animal and yeast genes. As the plant Golgi complex differs in many respects from its mammalian and yeast counterparts, the dbEST clones found can be directly used for various functional assays (immunoprecipitation, two-hybrid analysis, transgenic plants etc.) to test the exact roles of the encoded proteins and identify their functional partners, some of which may be specific for plants.
在酵母和哺乳动物细胞中已鉴定出许多参与内质网与高尔基体之间转运的蛋白质。目前，从两种主要的植物模式物种拟南芥和水稻中可获得大量部分cDNA序列，这使得鉴定来自非植物物种的已知基因/蛋白质的推定植物同源物成为可能。作者采用这种方法筛选表达序列标签数据库（dbEST），以检测参与内质网与高尔基体之间膜转运的蛋白质的植物同源物。这些部分序列的可用性将有助于筛选cDNA和基因组文库，否则这些文库的筛选是使用来自动物和酵母基因的异源探针进行的。由于植物高尔基体复合体在许多方面与其哺乳动物和酵母对应物不同，因此发现的dbEST克隆可直接用于各种功能测定（免疫沉淀、双杂交分析、转基因植物等），以测试编码蛋白质的确切作用并鉴定其功能伙伴，其中一些可能是植物特有的。
