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Molecular cloning, expression analyses of polymeric immunoglobulin receptor gene and its variants in grass carp (Ctenopharyngodon idellus) and binding assay of the recombinant immunoglobulin-like domains
- Source :
- Fishshellfish immunology. 88
- Publication Year :
- 2018
-
Abstract
- The Polymeric Immunoglobulin Receptor (pIgR) gene has been proved to play an important role in transporting polymeric immunoglobulin (Ig) in the mucosal tissues of mammals. pIgR gene also exists in teleost, but the genetic diversity and functions of this gene still need to be further explored. We obtained seven grass carp pIgR splicing transcripts, a full-length pIgR (CipIgR-1) and six truncated variants (CipIgR-2 to CipIgR-7). The full-length pIgR contained two immunoglobulin-like domains (ILD), a transmembrane domain (TMD) and a cytoplasmic domain (CyD). The CipIgR-2 lacked a small part in CyD, and CipIgR-3 lost TMD and CyD. Partial cDNA sequences of the other four grass carp pIgR variants (CipIgR-4 to CipIgR-7) were also cloned. The total expression levels of CipIgR and its variants in different tissues were detected by real-time quantitative PCR. The highest expression was found in the intestine, followed by the spleen and the skin. The function of the two extracellular ILDs of CipIgR was investigated based on its combining capacity with grass carp immunoglobulin M (IgM) and aquatic pathogenic bacteria. The cDNA sequences of two ILDs were cloned and expressed in Escherichia coli BL21 (DE3). Recombinant ILDs protein was purified and incubated with different bacteria respectively. Results of Western blot showed the recombinant protein could combine Bacillus subtilis, Vibrio parahaemolyticus, and Escherichia coli. In addition, binding activity of rILDs with grass carp IgM was detected. Collectively, these results indicated that multiple variants of pIgR gene in grass carp might be involved in the antibacterial immunity.
- Subjects :
- 0301 basic medicine
Carps
Aquatic Science
Molecular cloning
medicine.disease_cause
law.invention
03 medical and health sciences
Fish Diseases
law
Complementary DNA
medicine
Escherichia coli
Environmental Chemistry
Animals
Cloning, Molecular
Gene
Phylogeny
biology
Bacteria
Receptors, Polymeric Immunoglobulin
Genetic Variation
04 agricultural and veterinary sciences
General Medicine
biology.organism_classification
Molecular biology
Recombinant Proteins
Grass carp
030104 developmental biology
Immunoglobulin M
040102 fisheries
Recombinant DNA
biology.protein
0401 agriculture, forestry, and fisheries
Binding Sites, Antibody
Antibody
Immunoglobulin Domains
Polymeric immunoglobulin receptor
Subjects
Details
- ISSN :
- 10959947
- Volume :
- 88
- Database :
- OpenAIRE
- Journal :
- Fishshellfish immunology
- Accession number :
- edsair.doi.dedup.....c9ee148ffcead886f0db2a9c32d910a7