一株耐碱Mn-SOD高产细菌的分离、基因克隆及序列分析

首次从长白山温泉土中筛选得到了一株高产耐碱SOD的细菌，通过抑制剂试验确定了该菌所产SOD类型为Mn-SOD。通过形态特征、生理生化特征及16SrDNA基因序列分析，与芽孢杆菌Bacillus sp.MO6同源性高达100%，与地衣芽孢杆菌Bacillus licheniformis同源性为99%。根据地衣芽孢杆菌Mn-SOD序列，并结合GenBank中已发表的多种细菌Mn-SOD基因保守区，分别设计引物，PCR扩增获得600bp的Mn-SOD全基因序列和430bp的核心片段，克隆sodA全基因序列，构建重组质粒pMD18-SOD。

ISOLATION OF A BACTERIUM WITH HIGH PRODUCT OF ALKALINE MN-SUPEROXIDE DISMUTASE , GENECLONE AND SEQUENCE ANALYSIS

A high-product superoxide dismutase(SOD)bacterial strain was first obtained from the soil around the warm spring of Changbai Mountain,which is alkaline-resistant.The nature analysis by the inhibition of H2O2 and chloroform-ethanol indicated that the SOD was Mn-SOD.It belongs to bacillus genus by the modal character,physiological and biochemical character.The phylogenetic analyses based on 16SrDNA sequence indicated that the strain 110-2 was closed to Bacillus sp.MO6 with 100% identity and 99% with Bacillus licheniformis.According to the sodA sequence of Bacillus licheniformis and conservative regions of many kinds of bacillus,which were published in GenBank,two pairs of primers were designed and the complete sequence of Mn-SOD(600bp)and the core fragment(430bp)were amplified by PCR techniques.The recombinant plasmid pMD18-SOD was built successfully.