Heparin cofactor II (RbHCII) from rock bream (Oplegnathus fasciatus): Molecular characterization, cloning and expression analysis

Rock bream (Oplegnathus fasciatus) serpin, protease nexin-1: Transcriptional analysis and characterization of its antiprotease and anticoagulant activities

“…PN-1 homologues from various lineages indicate that the homology increases towards the C-terminus, along the SERPIN domain and reaches its maximum at the RCL region, similar to RbHCII. The mammalian PAI-1 members display closer homology to RbPN-1 than fish PAI-1 members which is in agreement with our previous findings of RbHCII (Umasuthan et al, 2011).…”

“…:*::** **:*:* greater degree of homology with the higher vertebrate PAI-1s than with the corresponding members from different lower vertebrates, including the fish (Table 2). Furthermore, matured HCII (Umasuthan et al, 2011) and RbPN-1 shared 23% of identity and 40% of similarity among them. ClustalW multiple alignment was employed to align complete AA sequences of RbPN-1 with PN-1 orthlogues of various vertebrates (Fig.…”

“…The immune challenge experiments were performed as described earlier (Umasuthan et al, 2011). For the injury experiment, a modified procedure from Wu et al (2004) was followed …”

“…The transcriptional analysis by qRT-PCR was performed as described earlier (Umasuthan et al, 2011). The RbPN-1, RbHCII gene-specific primers and ␤-actin primers (designed based on a Rb ␤-actin sequence; accession no.…”

A teleostean angiotensinogen from Oplegnathus fasciatus responses to immune and injury challenges

“…Table 1). Briefly, the reaction was performed in 20 mL mix containing 4 mL of diluted cDNA, 10 mL of 2 Â TaKaRa Ex TaqÔ SYBR premix, 0.5 mL of each primer (20 pmol/mL) and 5 mL dH 2 O using following cycle profile: one cycle of 95 C for 10 s, followed by 35 cycles of 95 C for 5 s, 58 C for 10 s and 72 C for 20 s and a final single cycle of 95 C for 15 s, 60 C for 30 s and 95 C for 15 s [22]. The relative RbAGT mRNA expression was determined by the Livak 2 ÀDDCT method [23], using b-actin as the reference gene.…”