中文摘要：

MAP激酶级联途径在信号传导过程中发挥着重要作用.以白菜型冬油菜＂陇油6号＂为材料,利用RT-PCR法克隆到全长MKK2基因cDNA,生物信息学分析表明油菜MKK2蛋白为亲水蛋白,包含有MAPKK蛋白所特有的磷酸化基序S/T-X_（3-5）-S/T.将其与大肠杆菌表达载体pET30a连接,构建原核表达载体pET30a-MKK2,并转化大肠杆菌BL21,经IPTG诱导表达后,SDS-PAGE以及Western blotting检测结果表明该基因表达了1个约43 kD的蛋白.实时荧光定量PCR结果显示该基因表达没有组织特异性,其转录受UV-B胁迫的诱导.

英文摘要：

MAP kinase cascade pathway plays an important role in the signal transduction process.MKK2 cDNA was cloned by RT-PCR from winter turnip rape（Brassica campestris L.） cultivar Longyou 6.Bioinformatic analysis showed that MKK2 protein is a hydrophilic protein,containing unique MAPKK protein phosphorylation motif S/TX_（3-5）-S/T.MKK2 cDNA was cloned into a pET30 a vector to construct the recombinant prokaryotic expression vector pET30a-MKK2.The pET30a-MKK2 was transformed to E.coli strain of BL21.After being induced by IPTG,a 43 kD recombinant protein was expressed and separated by SDS-PAGE electrophoresis and detected by western blotting.Real-time PCR results showed that there was nothing tissue-specific in the gene expression and transcription of MKK2 induced by UV-B stress.