中华绒螯蟹与长江华溪蟹胚胎发育相关miRNA识别与比较分析

Characterization and Comparative Analysis of MicroRNAs During Embryonic Development Between the Chinese Mitten Crab(Eriocheir sinensis) and the Freshwater Crab(Sinopotamon yangtsekiense)

利用Illumina/Solexa高通量测序技术,开展了中华绒螯蟹和长江华溪蟹胚胎的小RNA深度测序.对测序结果进行生物信息学分析,分别筛选获得中华绒螯蟹的8 569 743条和长江华溪蟹的8 719 465条干净序列(Clean Unique Reads),长度分布在20 nt^22 nt区间内的分别占有30.47%(Ejs-OB:中华绒螯蟹胚胎测序文库)和30.38%(SY-OJC:长江华溪蟹胚胎测序文库).用SOAP程序将小RNA定位到基因组,结果 Ejs-OB中分析筛选出5 745 279个小RNA,其中13 472种小RNA与基因组的序列匹配;SY-OJC中分析筛选出4 397 509个小RNA,其中18 407种小RNA与基因组的序列匹配.分类注释的结果显示,中华绒螯蟹和长江华溪蟹分别有6 293 445(Ejs-OB)和5 596 614(SY-OJC)条miRNA候选序列,但未预测发现新miRNA.在表达水平的差异分析结果中发现miR-1183、miR-1357、miR-1591、miR-2382的表达水平上调且差异显著.在两文库中搜寻到一些共同miRNA*序列,其中miR-139*、miR-1419g*、miR-1798*、miR-202*、miR-2068*和miR-454*也是表达变异较大.这些结果表明,miRNA可能在调控与中华绒螯蟹和长江华溪蟹胚胎发育相关的基因表达中起重要作用.

The deep sequencing of small RNA in Eriocheir sinensis and Sinopotamon yangtsekiense embryo samples was performed using the Illumina/Solexa high-throughput sequencing technologies. After analyzing the biological information of the sequencing results, 8 569 743 and 8 719 465 clean unique sequence data were screened in E. sinensis and S. yangtsekiense respectively, and the length distribution in the range of 20 n t -2 2 nt was 30.47% （ Ejs-OB ： S. embryonic sequencing library） and 30. 38% respectively （ SY-OJC ： 5. yangtsekiense embryonic sequencing library） . Mapping the small RNA tags to genome by SOAP to analyze their distribution on the genome,5 745 279 and 4 397 509 small RNAs of Ejs-OB and SY-OJC were screened respectively, 13 472 （ Ejs-OB ） and 18 407 （ SY-OJC ） of which were genome-matched reads. Annotated small RNA tags with different kinds of RNA, the results showed that a total of 6 293 445 and 5 596 614 miRNA candidate reads were searched in Ejs-OB and SY-OJC respectively. However,the novel miRNAs were not found. Expression profile analysis found that the expression level of miR-1183,miR-1357,miR-1591, miR-2382 were up-regulated and the difference were significant. Interestingly, some same miRNAs ＊ sequences were also searched in the two library, and miR-139＊ ,miR-1419g＊ ,miR-1798＊ ,miR-454＊ and miR-2068 ＊ of which were also have a significant differences in expression. These results indicated that these miRNAs may be involved in regulating gene expression associated with the development of the embryo in E. sinensis and 5. yangtsekiense.