title: Molecular cloning, sequencing and tissue-level expression of complement C3 of Labeo rohita (Hamilton, 1822).
abstract: Complement component C3 plays a central role in all known complement activation pathways. In the present study, we cloned, sequenced and analyzed the full-length cDNA sequence of Labeo rohita complement C3 (LRC3). The expression pattern of complement C3 mRNA in different tissues of healthy rohu and after challenge with Aeromonas hydrophila were evaluated using real-time PCR. The LRC3 cDNA sequence of rohu comprised of 5081 bp encoding a predicted protein of 1645 amino acids. The deduced amino acid sequence had the characteristic domain architecture. About eight domains specific to complement C3 are present in the sequence starting from signal peptide to netrin C345C (NTR) domain. The post-translational processing signal sequence (RKRR), the C3-convertase cleavage site sequence (LAR) and the canonical thiol-ester motif (GCGEQ) were found to be conserved in the LRC3. Real-time PCR analysis revealed the highest expression of C3 in liver and extra-hepatic expression of C3 was also observed in all the tissues studied. A. hydrophila challenge resulted in significant up-regulated expression of C3 transcripts in both liver and kidney at 6, 12, 24, 48 and 72 h post-infection.
authors.affiliation: Aquatic Environment and Health Management Division, Central Institute of Fisheries Education, Versova, Mumbai 400061, India.
authors.forename: K
authors.initials: K
authors.lastname: Pushpa
authors.affiliation: Fish Genetics and Biotechnology Division, Central Institute of Fisheries Education, Versova, Mumbai 400061, India.
authors.forename: P
authors.initials: P
authors.lastname: Gireesh-Babu
authors.affiliation: Aquatic Environment and Health Management Division, Central Institute of Fisheries Education, Versova, Mumbai 400061, India.
authors.forename: K V
authors.initials: KV
authors.lastname: Rajendran
authors.affiliation: Aquatic Environment and Health Management Division, Central Institute of Fisheries Education, Versova, Mumbai 400061, India.
authors.forename: C S
authors.initials: CS
authors.lastname: Purushothaman
authors.affiliation: Fish Nutrition, Biochemistry and Physiology Division, Central Institute of Fisheries Education, Kolkata Centre, Kolkata, West Bengal 700091, India.
authors.forename: S
authors.initials: S
authors.lastname: Dasgupta
authors.affiliation: Aquatic Environment and Health Management Division, Central Institute of Fisheries Education, Versova, Mumbai 400061, India. Electronic address: mmakesh@cife.edu.in.
authors.forename: M
authors.initials: M
authors.lastname: Makesh
mesh_terms: D016980:Aeromonas hydrophila / Q000502:physiology; D000595:Amino Acid Sequence; D000818:Animals; D003176:Complement C3 / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D003530:Cyprinidae; D018076:DNA, Complementary / Q000235:genetics / Q000378:metabolism; D005393:Fish Diseases / Q000235:genetics / Q000276:immunology* / Q000382:microbiology; D029941:Fish Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D016905:Gram-Negative Bacterial Infections / Q000235:genetics / Q000276:immunology / Q000382:microbiology / Q000662:veterinary*; D008969:Molecular Sequence Data; D010802:Phylogeny; D016415:Sequence Alignment / Q000662:veterinary
chemical_list: D003176:Complement C3; D018076:DNA, Complementary; D029941:Fish Proteins
keywords: Aeromonas hydrophila infection; Complement C3; Labeo rohita; Relative quantification; Rohu