Project description:The molecular mechanism involved in BmNPV resistance was investigated using a genome wide microarray in the midgut tissues of BmNPV infected resistant (Sarupat) and susceptible (CSR-2) Indian silkworm races. In the resistant race, 735 genes were upregulated and 589 genes were down regulated at 12 hours of BmNPV post infection. Similarly in case of susceptible race, 2183 genes were up regulated and 2115 genes were down regulated. Among the differentially expressed genes, nine upregulated and eight down regulated genes were validated using real time qPCR analysis. In Sarupat, the significantly upregulated genes are vacuolar protein sorting associated gene, X fin like protein and Carboxy peptidase E like protein in BmNPV infected midguts, whereas the prominent down regulated genes are glutamate receptor ionotropic kainite 2-like, BTB/POZ domain and transferrin. In the case of CSR-2, the considerably upregulated genes are Peptidoglycan recognition protein S6 precursor and rapamycin while the conspicuous down regulated genes are facilitated trehalose transporter and zinc transporter ZIP1-like gene. Our results provided a vital insights of Bombyx mori in reference with its molecular mechanism in immune response against the BmNPV invasion.
Project description:The molecular mechanism involved in BmNPV resistance was investigated using a genome wide microarray in the midgut tissues of BmNPV infected resistant (Sarupat) and susceptible (CSR-2) Indian silkworm races. In the resistant race, 735 genes were upregulated and 589 genes were down regulated at 12 hours of BmNPV post infection. Similarly in case of susceptible race, 2183 genes were up regulated and 2115 genes were down regulated. Among the differentially expressed genes, nine upregulated and eight down regulated genes were validated using real time qPCR analysis. In Sarupat, the significantly upregulated genes are vacuolar protein sorting associated gene, X fin like protein and Carboxy peptidase E like protein in BmNPV infected midguts, whereas the prominent down regulated genes are glutamate receptor ionotropic kainite 2-like, BTB/POZ domain and transferrin. In the case of CSR-2, the considerably upregulated genes are Peptidoglycan recognition protein S6 precursor and rapamycin while the conspicuous down regulated genes are facilitated trehalose transporter and zinc transporter ZIP1-like gene. Our results provided a vital insights of Bombyx mori in reference with its molecular mechanism in immune response against the BmNPV invasion. Organism : Bombyx mori , Agilent Custom Silkworm Gene Expression 4x44k Array (AMADID: 066047) designed by Genotypic Technology Private Limited.
Project description:Bombyx mori is one of the key lepidopteran model species, and is economically important for silk production and proteinaceous drug expression. Baculovirus and insect host are important natural biological models for studying host-pathogen interactions. The impact of Bombyx mori nucleopolyhedrovirus (BmNPV) infection on the proteome and acetylome of Bombyx mori ovarian (BmN) cells were explored to facilitate a better understanding of infection-driven interactions between BmNPV and host in vitro. The proteome and acetylome were profiled through 6-plex Tandem mass tag (TMT) labelling-based quantitative proteomics. Totally, 4,194 host proteins were quantified, of which 33 were up-regulated and 47 were down-regulated in BmN cells at 36 h post-infection. Based on the proteome, quantifiable differential Kac proteins were identified and functionally annotated to gene expression regulation, energy metabolism, substance synthesis and metabolism after BmNPV infection. Altogether, 644 Kac sites in 431 host proteins and 39 Kac sites in 22 viral proteins were identified and quantified in infected BmN cells. Our study demonstrated that BmNPV infection globally impacts the proteome and acetylome of BmN cells. The viral proteins are also acetylated by the host acetyltransferase. Protein acetylation is essential for cellular self-regulation and response to virus infection. This study provides new insights for understanding the host-virus interaction mechanisms, and the role of acetylation in BmN cellular response to viral infection.
Project description:Employed BmN cell after 10 MOI (multiplicity of infection) BmNPV 36 hpi (hour post-infection) to compare the proteomics by LC-MS/MS
Project description:Bombyx mori nucleopolyhedrovirus (BmNPV) is one of the most acute infectious diseases in silkworm, which has caused great economic loss in sericulture. Previous study showed that the content of components in mulberry leaves, particularly for moracin N, was increased after UV-B irradiation. In this study, the BmNPV resistance of silkworms reared on UV-B treated and moracin N spreaded mulberry leaves was improved. To uncover the mechanism of enhanced BmNPV resistance, silkworm midguts from UV-B treated mulberry leaves (BUM) and moracin N (BNM) groups were analyzed by SWATH-based proteomic technique. Of note, the abundance of ribosomal proteins in BUM and BNM groups was significantly changed to maintain the synthesis of total protein levels and cell survival. While, cytochrome c oxidase subunit II, calcium ATPase and programmed cell death 4 involved in apoptotic process were up-regulated in BNM group. Expressions of lipase-1, serine protease precursor, Rab1 protein, and histone genes were increased significantly in BNM group. These results suggest that moracin N might be the main active components in UV-B treated mulberry leaves to affect the BmNPV-resistance of silkworm, which could promote apoptotic cell death, enhance the organism immunity, and regulate the intercellular environment of cells in silkworm. It also presents an innovative process to reduce the mortality rate of silkworm infected with BmNPV.