根据基因库中细菌SOD的基因保守序列和Arthrobacter pascens DMDC12的N-末端氨基酸序列设计引物。采用PCR技术,以A.pascens DMDC12基因组为模板,克隆了A.pascens DMDC12中Mn—SOD的567bp的基因片段;再结合该基因片段和A.pascens DMDC12中SOD的N-末端氨基酸序列的有关信息,设计包含该sod完整ORF区域的引物,成功扩增了A.pascens DMDC12中Mn—SOD的基因序列,新sod序列(sodAP)已提交Gen-Bank(收录号DQ779150)。利用生物学软件对该序列进行分析表明,该sodAP序列的ORF区域全长651bp,编码216个氨基酸;该序列和Arthrobactersp.FB24的SOD基因序列同源性为86%。
A primer was designed according to conservative sequence of SOD gene in GenBank and amino acid sequence of N-terminal in Arthrobacter pascerts (Ap) DMDC12, then a 567 bp gene fragment of Mn-SOD from Ap DMDC12 was cloned adopting PCR and using genome of AP DMDC12 as a template. Combining the gene fragment and relevant information of N-terminal amino acid sequence of SOD from Ap DMDC12 a primer containing SOD complete ORF domain was designed and successfully amplified the Mn-SOD gene sequence in Ap DMDCI2. The new SOD sequence (SODAP) was submitted to GenBank (No. DQ779150). Sequence analysis using biological software indicated that the length of ORF domain of SODAP was 651 bp encoding 216 amino acids, the homology of the sequence was 86% with SOD sequence from Arthrobacter sp. FB24.