中文摘要：

将拟南芥α-dioxygenase2（DOX2）基因编码区克隆到原核表达载体pGEX-5X-1获得重组表达载体pGEX-DOX2,转化至大肠杆菌（Escherichia coli）BL21（DE3）pLysS和BL21（DE3）-RIPL codon＋菌株中进行诱导表达。SDS-PAGE和Western blot结果表明,融合蛋白在BL21（DE3）-RIPL codon＋中得到了高效表达,融合蛋白表观分子量约为98kD,占菌体总蛋白的38%。经GST亲和柱层析进一步纯化获得了可溶性重组蛋白;愈创木酚法测试表明,可溶性重组蛋白不具有过氧化物酶活性。利用Rare codon Caltor进行的密码子偏爱性分析显示,该基因读码框中含有88个大肠杆菌稀有密码子,占总密码子（631个）的14%;SignalP、CELLOv.2.5、PSLpred和Softberry-ProtComp Version6.1等软件分析结果显示,拟南芥DOX2可能定位于细胞质。

英文摘要：

The encoding sequence of Arabidopsis α-dioxygenase 2 （DOX2） gene was inserted into pGEX-5X-1 to obtain pGEX-DOX2. The recombinant vector was transformed into Escherichia. coli BL21 （ DE3 ）pLysS and BL21 （DE3）-RIPL codon^＋, respectively. A fusion protein about 98 kD was detected by SDS-PAGE and Western blot in the induced recombinant BL21 （DE3）-RIPL codon^＋ strain, and it accounts for 38% of the total bacterial proteins. The target protein was purified by the GST chromatography column, and the peroxidase activity of the purified protein was tested using the guaiacol method, the result showed that the soluble fusion protein had no detectable peroxidase activity. Using rare codon Caltor software, 88 rare codons of E.coli were found in the DOX2 ORF. The subcellular localization analysis result predicted by Signal P, CELLO v.2.5, PSLpred and Softberry-ProtComp Version 6.1 indicated that the Arabidopsis DOX2 located in cytoplasm.