光照与温度对拟南芥tak基因表达与LHC磷酸化的影响  

Effects of Light and Temperature on The Expression of tak Gene and Phosphorylation of LHC in Arabidopsis thaliana

作　　者：蒋佳宏[1] 王东[1] 胡源[1] 高秀[1] 杜林方[1,2] 

机构地区：[1]四川大学生命科学学院,生物资源与生态环境教育部重点实验室,成都610064 [2]四川大学纳米生物医学技术与膜生物学研究所,成都610041

出　　处：《生物化学与生物物理进展》2009年第9期1202-1207,共6页Progress In Biochemistry and Biophysics

基　　金：国家自然科学基金资助项目(30870181)~~

摘　　要：TAK1和TAK2是拟南芥核基因编码的蛋白激酶,TAK1可能参与植物状态转换中LHCⅡ蛋白磷酸化的调控.根据TAK1蛋白序列,合成了一段含12个氨基酸残基的亲水短肽,与牛血清白蛋白(BSA)偶联并免疫家兔得到TAK1抗体.经免疫双扩散和蛋白质印迹检测,该多肽抗体效价和特异性较高.借助TAK1抗体以及磷酸化抗体进行杂交试验,结合特异引物进行半定量PCR,研究了光照与温度对拟南芥tak1和tak2基因表达的影响.结果表明,光强与温度调节LHCⅡ蛋白磷酸化,也会在转录水平和蛋白质水平上影响tak1和tak2基因表达.LHCⅡ蛋白磷酸化对光响应趋势与TAK1蛋白水平变化相同.此外,低光照促进tak2基因表达,tak2基因转录表达受温度的影响较小.tak1和tak2基因表达调控方式不同,可能与启动子响应元件的不同相关.TAK1 and TAK2 are both nuclear-encoded kinases. TAK1 may be involved in the phosphorylation of LHC Ⅱ in state transitions. After the analysis of conservative activity regions on TAK1 and TAK2 sequences, a segment of hydrophilic polypeptide consisting of 12 amino acid residues was devised and linked to bovine serum albumin （BSA） before injection to the rabbit. The polyclonal antibody against TAK1 raised was examined by agar gel immunodiffusion （AGID） test and Western blot analysis, then it was used to study the expression of tak1 and tak2 influenced by the change of light and temperature in RT-PCR with specific primers and the possible regulation relationship between TAK1 and LHC Ⅱ phosphorylation especially by using TAK1 serum and P-Thr antibody. The result indicated that light and temperature regulated the phosphorylation of LHC Ⅱ and took effect on the transcription and translation level of tak1 and tak2, but the response of LHC Ⅱ phosphorylation to light did not coincided with the quantity of TAK1 protein. Furthermore, low light density could enhance the expression of tak2, but temperature impacted tak2 little. The different regulation pattern of takl and tak2 may derive from the different elements of promoter.

关 键 词：TAK1 TAK2 LHC Ⅱ磷酸化 合成多肽 基因表达 光照与温度影响 

分 类 号：Q5[生物学—生物化学]