家蚕新的非编码RNA功能初探

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.

Functional Dissection of Novel Non-coding RNA in Silkworm Bombyx mori

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 263 bp upstream of Bm-86,which indicated that Bm-86 might be transcribed independently.Further analysis showed that the expression of Bm-86 exhibited negative correlation with its host gene eIF5A,notably in silkgland,which indicated that Bm-86 might be involved in the development of silkgland through regulating the expression of eIF5A.