应用mRNA差异显示技术筛选磁螺菌MSR-1磁小体合成相关基因

Screening of the Genes Related to the Synthesis of Magnetosome of Magnetic Spirillum MSR-1 by m RNA differential Display Technology

为寻找Magnetospirillum gryphiswaldense MSR-1中与磁小体合成相关的基因,实验利用氧气浓度对MSR-1生长的影响,应用m RNA差异显示技术筛选与磁小体合成相关的c DNA片段,经测序后获得7条差异片段序列.共得到M-1,M-2,…,M-7共7条差异片段,然后进行生物信息学分析.片段M-2和片段M-3推测蛋白质均含有保守结构域Mqs R,根据Mqs R对生物膜形成有重要调节作用推测此片段可能与磁小体外膜的形成有关.其中M-4推测蛋白质具有保守结构域rpo H2,推测此片段编码的蛋白质可能是识别MSR-1中磁小体合成相关基因起始位点的酶.片段M-5推测的蛋白质具有PPK2超家族和ALDH-SF超家族,推测片段M-5编码蛋白质的功能可能是为磁小体提供能量.片段M-6,M-7编码蛋白质推测功能可能也是为MSR-1合成磁小体提供能量.

To search for genes related to magnetosome synthesis in Magnetospirillum gryphiswaldense MSR-1,in this experiment,we made use of the effect of oxygen concentration on MSR-1 growth and screened the correlated cDNA fragments using mRNA differential display PCR. After sequencing,7 differential fragments were identified,namely M-1,M-2,…,M-6 and M-7. These were then analyzed bioinformatically. The frag？ments M-2 and M-3 encoded proteins with conserved domain MqsR. Due to the important role of MqsR in regulating the formation of biological membranes,we predicted that these two proteins may be involved in the formation of the outer membrane of magnetosome. M-4 encoded protein with conserved domain rpoH2 and this protein was probably an enzyme for recognizing the start site of the genes correlated with magnetosome synthesis. Protein encoded by M-5 contained PPK2 superfamily and ALDH-SF superfamily and was predict？ed to function as energy provider for magnetosome. Proteins encoded by M-6 and M-7 were also predicted to provide energy for magnetosome synthesis.