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Alternative splicing and trans-splicing events revealed by analysis of the Bombyx mori transcriptome

机译:通过家蚕转录组的分析揭示了可变的剪接和反剪接事件

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Alternative splicing and trans-splicing events have not been systematically studied in the silkworm Bombyx mori. Here, the silkworm transcriptome was analyzed by RNA-seq. We identified 320 novel genes, modified 1140 gene models, and found thousands of alternative splicing and 58 trans-splicing events. Studies of three SR proteins show that both their alternative splicing patterns and mRNA products are conserved from insect to human, and one isoform of Srsf6 with a retained intron is expressed sex-specifically in silkworm gonads. Trans-splicing of mod(mdg4) in silkworm was experimentally confirmed. We identified integrations from a common 5′-gene with 46 newly identified alternative 3′-exons that are located on both DNA strands over a 500-kb region. Other trans-splicing events in B. mori were predicted by bioinformatic analysis, in which 12 events were confirmed by RT-PCR, six events were further validated by chimeric SNPs, and two events were confirmed by allele-specific RT-PCR in F 1 hybrids from distinct silkworm lines of JS and L10, indicating that trans-splicing is more widespread in insects than previously thought. Analysis of the B. mori transcriptome by RNA-seq provides valuable information of regulatory alternative splicing events. The conservation of splicing events across species and newly identified trans-splicing events suggest that B. mori is a good model for future studies. Published by Cold Spring Harbor Laboratory Press.
机译:蚕Bombyx mori中尚未系统地研究其他剪接和转拼事件。在此,通过RNA-seq分析家蚕转录组。我们鉴定了320个新基因,修改了1140个基因模型,发现了成千上万个可变剪接和58个反式剪接事件。对三种SR蛋白的研究表明,它们的选择性剪接模式和mRNA产物在昆虫与人之间均是保守的,并且具有保留内含子的Srsf6的一种同工型在蚕的性腺中性别特异性地表达。实验证实了mod(mdg4)在蚕中的转拼。我们从一个共同的5'基因与46个新近鉴定的替代3'外显子(位于500 kb区域的两条DNA链上)中鉴定出整合。通过生物信息学分析预测了家蚕中的其他反式剪接事件,其中RT-PCR证实了12个事件,嵌合SNP进一步证实了6个事件,等位基因特异性RT-PCR在F 1中证实了两个事件JS和L10不同家蚕品系的杂种,表明在昆虫中反式剪接比以前认为的要广泛。通过RNA-seq对桑蚕双歧杆菌转录组的分析提供了调控性选择性剪接事件的有价值的信息。跨物种的剪接事件和新发现的跨剪接事件的保守性表明,桑蚕芽孢杆菌是未来研究的良好模型。由冷泉港实验室出版社出版。

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