用马氏珠母贝基因组序列,通过生物信息学方法获得Pm-miR-183的前体,长度76 bp,具有典型的茎环结构.成熟序列及前体序列的同源性分析均显示较高的种间保守性.Pm-miR-183前体序列(Pm-pre-miR-183)的系统进化树分析,Pm-pre-miR-183与帽贝同源性较高.靶向关系分析,Pm-miR-183与矿化基因及免疫相关基因均存在靶向作用位点.实时荧光定量分析,Pm-miR-183在马氏珠母贝的各组织中均有表达.注射 Pm-miR-183 mimics进行过表达,Pm-miR-183的表达量在套膜区(MP)中显著上调(P<0.05).扫描电子显微镜(scanning electron microscope,SEM)检测,Pm-miR-183过表达会导致贝壳珍珠层生长出现紊乱.%MiRNAs are non-coding small RNAs involved in negatively transcriptional regulation. In our previous study, we had identified a miR-183 homologue in Pinctada fucata martensii by Solexa deep sequencing.Based on bioinformatics results obtained from whole genome sequence of P. f.martensii, the precursor Pm-miR-183(Pm-pre-miR-183)obtained has a length of 76 bp and a typical stem-loop structure. Homology analysis of mature sequence and previous sequence indicated that they were highly conserved with other species. Phylogenetic analysis of precursor sequence showed that Pm-pre-miR-183 had a higher homology to Lottia gigante. The potential interactions among Pm-miR-183 and several mineralized genes and immune-related genes were obtained by target prediction. Pm-miR-183 was expressed in different tissues. In addition, after injection of Pm-miR-183 mimics, the expression of Pm-miR-183 was significantly up-regulated in the mantle pallial. The result of SEM showed that there were adnormal growth of nacre in shell. This study shows that Pm-miR-183 could be involved in the formation of pearl oyster in P.f.martensii.
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