水稻蛋白激酶的规模化克隆、表达及活性研究(简报)

HIGH THROUGHPUT CLONING,EXPRESSION AND AUTOPHOSPHORYLATION ANALYSIS OF RICE KINASES

蛋白激酶几乎与所有重要的发育代谢过程有关，已知在植物的发育、自交不亲和、雄性不育、抗逆和抗病等生命活动过程中起重要的调控作用。蛋白激酶在各种生物中广泛存在，根据网络公布的水稻全基因组序列图谱，通过同源序列比对．TIGR Rice Genome Annotation—Release4共找到了1532个具有激酶结构域的蛋白质（PF00069）.

Protein kinases are essential for the regulation of cell growth and development in plant. Based on rice genomic sequences obtained from the database, bioinformatics analysis of homology sequences suggested over 1500 kinases in rice. In this study, 32 PCR primers were synthesized based on conserved domains of known kinases, 30 intron-free kinase PCR products were obtained, among them, 27 kinases were expressed in Saccharomyces cerevisiae with visible band after separated by SDS-PAGE and stained by Coomassie Blue. According to annotation, 27 kinases belong to 7. groups in the first class on PlantsP （http：//plantsp.genomics.purdue.edu/）, Receptor like cytoplasmic kinase Ⅶ, CRPK1 Like Kinase, Crinkly 4_Like kinase, Plant External Response Like Kinase, S-Domain Kinase （Type 2）, legume Lectin Domain Kinase, Wall associate kinase, respectively. Furthermore, 17 kinases showed enzyme activity detected by autophosphorylation experiment. The results provide critical materials to the study of biochemical prosperities and substrate identification.