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Cloning, characterization and expression analysis of coagulation factor II gene in grass carp (Ctenopharyngodon idella).
- Source :
-
Genetics and molecular research : GMR [Genet Mol Res] 2015 Oct 29; Vol. 14 (4), pp. 13764-78. Date of Electronic Publication: 2015 Oct 29. - Publication Year :
- 2015
-
Abstract
- Here, we characterized the structure and function of the coagulation factor II (FII) gene in grass carp and determined its role in coagulation mechanisms. The FII gene EST was obtained using a constructed splenic transcriptome database; the full-length FII gene sequence was obtained by 3' and 5' RACE. The open reading frame (ORF) of FII was cloned and the full-length gene was found to be 1718 bp, with an ORF of 1572 bp; the gene contained a 25 bp 5'-untranslated region (UTR) and 108 bp 3'-UTR. The ORF encoded 524 amino acids, including 74 alkaline amino acids (arginine and lysine) and 69 acidic amino acids (aspartic acid and glutamic acid). The theoretical pI was 6.22. The calculated instability index (II) was 39.81, indicating that FII was a stable protein; the half-life period was predicted to be approximately 30 h. Amino acid sequence comparisons indicated that grass carp FII showed most similarity (71%) to FII of Takifugu rubripes, followed by Oplegnathus fasciatus (48% similarity) and Larimichthys crocea (47% similarity). A real-time reverse transcription PCR analysis showed that under normal circumstances, FII was most highly expressed in the liver, followed by the gill, spleen, thymus, and head-kidney (P < 0.001). After injection of the grass carp reovirus 873 (GCRV873), the pattern of FII expression was significantly altered (P < 0.001); gene expression was high after injection, suggesting a response involving the initiation of the coagulation system and defense of the body in combination with the platelet and complement system.
- Subjects :
- Amino Acid Sequence
Animals
Base Sequence
DNA, Complementary
Evolution, Molecular
Models, Molecular
Molecular Sequence Data
Organ Specificity genetics
Phylogeny
Protein Conformation
Prothrombin chemistry
RNA Splicing
RNA, Messenger genetics
Sequence Alignment
Sequence Analysis, DNA
Carps genetics
Cloning, Molecular
Gene Expression
Prothrombin genetics
Subjects
Details
- Language :
- English
- ISSN :
- 1676-5680
- Volume :
- 14
- Issue :
- 4
- Database :
- MEDLINE
- Journal :
- Genetics and molecular research : GMR
- Publication Type :
- Academic Journal
- Accession number :
- 26535692
- Full Text :
- https://doi.org/10.4238/2015.October.28.39