家蚕新的非编码RNA功能初探
Functional Dissection of Novel Non-coding RNA in Silkworm Bombyx mori
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摘要: 构建了家蚕50～500 nt非编码RNA的cDNA文库,发现了189个新的ncRNA,其中一个小RNA-Bm-86在家蚕幼虫和蛹中的表达量显著高于卵和成虫.利用生物信息学软件对该ncRNA的特性及其与上下游基因的关系进行了深入分析,并利用Northern杂交和半定量RT-PCR技术对该ncRNA与其宿主基因在家蚕不同组织的表达情况进行了研究.结果发现,该ncRNA属于H/ACA box snoRNA家族,是由家蚕eIF5A基因的第二个内含子转录产生的,且在家蚕中没有明显的靶标位点.分析其上下游基因结构发现,在其上游263 bp处存在着一个可能的启动子位点,表明其有独立转录的倾向.进一步分析Bm-86与其宿主基因eIF5A在家蚕不同组织部位的表达情况发现,二者在表达上存在着负相关的关系,且该现象在丝腺中尤其明显.推测Bm-86可能通过影响eIF5A基因的表达参与到家蚕丝腺发育调控过程中Abstract: The cDNA library of 50-500 nt ncRNAs in Bombyx mori were constructed,and a small RNA Bm-86 accumulated more in larva and pupa during development was found.To further dissect the function of this ncRNA,bioinformatics analysis,northern blot and RT-PCR were taken out.Results showed that Bm-86 was an H/ACA box small nucleolar RNA,located in the second intron of eIF5A,with no predicted pseudouridylation modification site in rRNAs,snRNAs and tRNAs of B.mori.Moreover,a putative promoter were located 26...
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