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Molecular characterization of a peritrophic membrane protein from the silkworm, Bombyx mori
Xiaolong Hu1, Lin Chen, Rui Yang
1College of Animal Sciences, Zhejiang University, Hangzhou, 310058, China.
Abstract:
The peritrophic membrane lines the gut of most insects at one or more stages of their life cycles. It facilitates the digestive processes in the guts and protects from invasion by pathogens or food particles. In the current study, a novel PM protein, designated as BmMtch, was identified from the silkworm, Bombyx mori. The open reading frame of BmMtch is 888 bp in length, encoding 295 amino acid residues consisting of two domains (Mito_carr domains) and three transmembrane regions. They are localized on the 11th chromosome as single copy with one exon only. Quantitative real time PCR analysis (qRT-PCR) revealed that BmMtch was mainly expressed in larval fat bodies, Malpighian tubules, testis and ovaries, and could be detected through all stages of the life cycle of silkworm. Immuno-fluorescence analysis indicated that BmMtch was localized within the goblet cell of larval midgut. Western blotting analysis showed that BmMtch were detected in total proteins of PM and larval midgut. The characteristics of BmMtch indicated that BmMtch represents a novel member of insect PM proteins, without chitin-binding domains.
Insights
Researchers identified a novel peritrophic membrane (PM) protein, BmMtch, in the silkworm Bombyx mori. This protein plays a role in gut function and protection, offering new insights into insect physiology.
Area of Science:
- Insect molecular biology
- Gastrointestinal physiology
- Biochemistry
Background:
- The peritrophic membrane (PM) is crucial for insect gut function, aiding digestion and defense.
- Understanding PM composition is key to elucidating insect physiology and developing pest control strategies.
Purpose of the Study:
- To identify and characterize a novel protein component of the peritrophic membrane in the silkworm, Bombyx mori.
- To investigate the expression patterns and localization of this new protein.
Main Methods:
- Bioinformatic analysis of the silkworm genome to identify potential PM proteins.
- Gene cloning and sequencing of the novel protein, BmMtch.
- Quantitative real-time PCR (qRT-PCR) for expression analysis.
- Immunofluorescence and Western blotting for protein localization and detection.
Main Results:
- A novel PM protein, BmMtch, was identified in Bombyx mori, featuring two Mito_carr domains and three transmembrane regions.
- BmMtch is encoded by a single exon on chromosome 11 and expressed in larval fat bodies, Malpighian tubules, and reproductive organs across all life stages.
- Immunofluorescence confirmed BmMtch localization in larval midgut goblet cells, and Western blotting detected it in PM and midgut proteins.
Conclusions:
- BmMtch represents a novel class of insect PM proteins, distinct from those with chitin-binding domains.
- Its presence in various tissues and life stages suggests a significant role in silkworm gut physiology and development.
