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Molecular cloning, characterization and expression of the complement component Bf/C2 gene in grass carp
Yubang Shen1, Junbin Zhang, Xiaoyan Xu
1Key Laboratory of Exploration and Utilization of Aquatic Genetic Resources, Shanghai Ocean University, Ministry of Education, Shanghai Ocean University, Shanghai, PR China.
Fish & Shellfish Immunology
|February 28, 2012
Summary
Researchers characterized two grass carp genes, gcBf/C2A and gcBf/C2B, crucial for the complement system. These genes showed distinct expression patterns in response to infection and development, enhancing our understanding of fish immunity.
Area of Science:
- Immunology
- Molecular Biology
- Fish Science
Background:
- The complement system is vital for host defense, bridging innate and acquired immunity.
- Factor B (Bf) and C2 are key components of complement pathways, specifically the alternative and classical pathways, respectively.
Purpose of the Study:
- To clone and characterize two grass carp Bf/C2 genes, gcBf/C2A and gcBf/C2B.
- To investigate the expression patterns of these genes in various tissues and developmental stages.
- To analyze the regulation of gcBf/C2A and gcBf/C2B in response to bacterial challenge.
Main Methods:
- Gene cloning and sequencing of grass carp gcBf/C2A and gcBf/C2B.
- Bioinformatic analysis of gene and protein sequences, including comparison with human and mouse homologs.
- Quantitative real-time PCR (qPCR) to assess gene expression in different tissues, developmental stages, and after Aeromonas hydrophila infection.
Main Results:
- Two grass carp Bf/C2 genes, gcBf/C2A (2259 bp) and gcBf/C2B (3004 bp), were cloned and characterized.
- Amino acid identity between gcBf/C2A and gcBf/C2B was 30.7%, with lower identity to human and mouse Bf/C2 genes.
- Both genes were widely expressed, with highest levels in the liver. gcBf/C2A upregulated post-infection, while gcBf/C2B showed significant upregulation in head kidney. Developmental expression peaked early post-hatching.
Conclusions:
- The study provides a comprehensive characterization of grass carp Bf/C2 genes.
- Expression analysis reveals tissue-specific and developmental regulation of these complement components.
- Findings contribute to understanding the role of the complement system in fish immunity and disease resistance.
