title: Cloning, characterization and expression analysis of coagulation factor II gene in grass carp (Ctenopharyngodon idella).
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.
authors.affiliation: College of Animal Science and Technology, Hunan Agricultural University, Changsha, China.
authors.forename: B H
authors.initials: BH
authors.lastname: Xu
authors.affiliation: College of Animal Science and Technology, Hunan Agricultural University, Changsha, China.
authors.forename: K J
authors.initials: KJ
authors.lastname: Chen
authors.affiliation: College of Animal Science and Technology, Hunan Agricultural University, Changsha, China.
authors.forename: Y B
authors.initials: YB
authors.lastname: Yao
authors.affiliation: College of Animal Science and Technology, Hunan Agricultural University, Changsha, China.
authors.forename: Q L
authors.initials: QL
authors.lastname: Liu
authors.affiliation: College of Animal Science and Technology, Hunan Agricultural University, Changsha, China xiaotiaoyi605@163.com.
authors.forename: T Y
authors.initials: TY
authors.lastname: Xiao
authors.affiliation: College of Animal Veterinary and Medicine, Hunan Agricultural University, Changsha, China.
authors.forename: J M
authors.initials: JM
authors.lastname: Su
authors.affiliation: College of Animal Science and Technology, Hunan Agricultural University, Changsha, China.
authors.forename: H Z
authors.initials: HZ
authors.lastname: Peng
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002347:Carps / Q000235:genetics*; D003001:Cloning, Molecular; D018076:DNA, Complementary; D019143:Evolution, Molecular; D015870:Gene Expression; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009928:Organ Specificity / Q000235:genetics; D010802:Phylogeny; D011487:Protein Conformation; D011516:Prothrombin / Q000737:chemistry / Q000235:genetics*; D012326:RNA Splicing; D012333:RNA, Messenger / Q000235:genetics; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA
chemical_list: D018076:DNA, Complementary; D012333:RNA, Messenger; D011516:Prothrombin