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菲律宾饲养的桑蚕丝腺中基因表达的全球分析

Global Profiling of Genes Expressed in the Silk Glands of Philippine-Reared Mulberry Silkworms .

作者信息

de la Peña Pauline Nicole O, Lao Adria Gabrielle D, Bautista Ma Anita M

机构信息

National Institute of Molecular Biology and Biotechnology, University of the Philippines Diliman, Quezon City 1101, Philippines.

出版信息

Insects. 2022 Jul 24;13(8):669. doi: 10.3390/insects13080669.

Abstract

RNA sequencing was used to assemble transcriptome data for Philippine-reared silkworm and compare gene expression profiles of strains reared in high- and low-temperature environments. RNA was isolated from the silk glands of fifth instar larvae and mRNA-enriched libraries were sequenced using Illumina NextSeq 500. Transcriptome reads were assembled using reference-based and de novo assemblers, and assemblies were evaluated using different metrics for transcriptome quality, including the read mapping rate, E90N50, RSEM-eval, and the presence of single-copy orthologs. All transcriptome assemblies were able to reconstruct >40,000 transcripts. Differential expression analysis found 476 differentially expressed genes (DEGs; 222 upregulated, 254 downregulated) in strains reared in different temperatures. Among the top DEGs were myrosinase, heat shock proteins, serine protease inhibitors, dehydrogenases, and regulators of the juvenile hormone. Validation of some of the top DEGs using qPCR supported the findings of the in silico analysis. GO term enrichment analysis reveals an overrepresentation of GO terms related to nucleotide metabolism and biosynthesis, lipid and carbohydrate metabolic processes, regulation of transcription, nucleotide binding, protein binding, metal binding, catalytic activity, oxidoreductase activity, and hydrolase activity. The data provided here will serve as a resource for improving local strains and increasing silk production of Philippine-reared B. mori strains.

摘要

RNA测序用于组装菲律宾饲养家蚕的转录组数据,并比较在高温和低温环境中饲养的品系的基因表达谱。从五龄幼虫的丝腺中分离RNA,并使用Illumina NextSeq 500对富含mRNA的文库进行测序。使用基于参考和从头组装器对转录组读数进行组装,并使用不同的转录组质量指标对组装进行评估,包括读数映射率、E90N50、RSEM-eval和单拷贝直系同源物的存在。所有转录组组装都能够重建超过40,000个转录本。差异表达分析发现,在不同温度下饲养的品系中有476个差异表达基因(DEG;222个上调,254个下调)。在顶级DEG中,有黑芥子酶、热休克蛋白、丝氨酸蛋白酶抑制剂、脱氢酶和保幼激素调节剂。使用qPCR对一些顶级DEG进行验证支持了计算机分析的结果。GO术语富集分析揭示了与核苷酸代谢和生物合成、脂质和碳水化合物代谢过程、转录调控、核苷酸结合、蛋白质结合、金属结合、催化活性、氧化还原酶活性和水解酶活性相关的GO术语的过度表达。此处提供的数据将作为改善本地品系和提高菲律宾饲养的家蚕品系丝绸产量的资源。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9589/9329738/8856b5940f15/insects-13-00669-g002.jpg

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