Division of Fisheries Biotechnology & Molecular Biology, Department of Biotechnology, Faculty of Science and Humanities, SRM University, Kattankulathur 603 203, Chennai, Tamil Nadu, India; Department of Microbiology, SRM Arts & Science College, Kattankulathur 603 203, Chennai, India.
Division of Fisheries Biotechnology & Molecular Biology, Department of Biotechnology, Faculty of Science and Humanities, SRM University, Kattankulathur 603 203, Chennai, Tamil Nadu, India.
Fish Shellfish Immunol. 2016 Aug;55:149-58. doi: 10.1016/j.fsi.2016.05.030. Epub 2016 May 24.
A transcriptome wide analysis of the constructed cDNA library of snakehead murrel Channa striatus revealed a full length cDNA sequence of coagulation factor X. Sequence analysis of C. striatus coagulation factor X (CsFX) showed that the cDNA contained 1232 base pairs (bp) comprising 1209 bp open reading frame (ORF). The ORF region encodes 424 amino acids with a molecular mass of 59 kDa. The polypeptide contains γ-carboxyglutamic acid (GLA) rich domain and two epidermal growth factor (EGF) like domains including EGF-CA domain and serine proteases trypsin signature profile. CsFX exhibited the maximum similarity with fish species such as Stegastes partitus (78%), Poecilia formosa (76%) and Cynoglossus semilaevis (74%). Phylogenetically, CsFX is clustered together with the fish group belonging to Actinopterygii. Secondary structure of factor X includes alpha helix 28.54%, extended strand 20.75%, beta turn 7.78% and random coil 42.92%. A predicted 3D model of CsFX revealed a short α-helix and a Ca(2+) (Gla domain) binding site in the coil. Four disulfide bridges were found in serine protease trypsin profile. Obviously, the highest gene expression (P < 0.05) was noticed in blood. Further, the changes in expression of CsFX was observed after inducing with bacterial (Aeromonas hydrophila) and fungal (Aphanomyces invadans) infections and other synthetic immune stimulants. Variation in blood clotting time (CT), prothrombin time (PT) and activated prothromboplastin time (APTT) was analyzed and compared between healthy and bacterial infected fishes. During infection, PT and APTT showed a declined clotting time due to the raised level of thrombocytes.
通过对蛇头鱼 Channa striatus 构建的 cDNA 文库进行转录组全分析，揭示了凝血因子 X 的全长 cDNA 序列。序列分析表明，蛇头鱼凝血因子 X (CsFX) cDNA 包含 1232 个碱基对 (bp)，其中包括 1209bp 的开放阅读框 (ORF)。ORF 区编码 424 个氨基酸，分子量为 59kDa。该多肽含有 γ-羧基谷氨酸 (GLA) 丰富结构域和两个表皮生长因子 (EGF) 样结构域，包括 EGF-CA 结构域和丝氨酸蛋白酶胰蛋白酶特征结构域。CsFX 与 Stegastes partitus（78%）、Poecilia formosa（76%）和 Cynoglossus semilaevis（74%）等鱼类物种具有最大的相似性。系统发育分析表明，CsFX 与 Actinopterygii 鱼类群聚在一起。凝血因子 X 的二级结构包括α螺旋 28.54%、延伸链 20.75%、β转角 7.78%和无规卷曲 42.92%。CsFX 的预测 3D 模型显示了一个短的α-螺旋和一个 Ca(2+)（Gla 结构域）结合点在卷曲中。在丝氨酸蛋白酶胰蛋白酶特征结构域中发现了四个二硫键。显然，在血液中观察到最高的基因表达 (P<0.05)。此外，在细菌（嗜水气单胞菌）和真菌（阿氏单极虫）感染以及其他合成免疫刺激物诱导后，观察到 CsFX 的表达变化。分析并比较了健康和细菌感染鱼类之间的凝血时间 (CT)、凝血酶原时间 (PT) 和活化部分凝血活酶时间 (APTT) 的变化。在感染过程中，由于血小板水平升高，PT 和 APTT 显示出凝血时间缩短。
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