A C1 inhibitor ortholog from rock bream (Oplegnathus fasciatus): Molecular perspectives of a central regulator in terms of its genomic arrangement, transcriptional profiles and anti-protease activities of recombinant peptide

Molecular characterization and comparative expression analysis of two teleostean pro-inflammatory cytokines, IL-1β and IL-8, from Sebastes schlegeli

“…A series of PCRs were carried out with sequence specific primer pairs (Table 1) in order to identify clones containing full-length RfIL-1β and RfIL-8 genomic sequences according to the manufacturer's instructions. Positive clones were located as described elsewhere (Umasuthan et al, 2014). Two individually identified BAC clones, one containing a full-length RfIL-1β and one containing a full-length RfIL-8 gene, were sequenced by GS-FLX™ system (Macrogen, Korea).…”

Two NF-κB inhibitor-alpha (IκBα) genes from rock bream (Oplegnathus fasciatus): Molecular characterization, genomic organization and mRNA expression analysis after immune stimulation

“…A random shear bacterial artificial chromosome (BAC) library of rock bream genomic DNA was custom constructed (Lucigen, USA) and used to screen OfIkBa-A and OfIkBa-B genes, following pooling and super pooling strategies, and performing polymerase chain reaction (PCR), using the sequence specific primer pairs (OfIkBa-A: forward, 5 0 -TAACCTGCTGTGCTTCAGCGTGAT-3 0 and reverse, 5 0 -ATCCACCATGTTCTCCACCAGTGA-3 0 and OfIkBa-B: forward, 5 0 -ATCAATGCACAGGAACAGTGCAGC-3 0 and reverse 5 0 -AAGCAGTTAA-CATCAGCGCCAAGG-3 0 ) as described previously [24,25]. BAC DNA from positive clones were isolated and purified using QIAGEN Large-Construct Kit, and were subjected to sequencing (Macrogen, Republic of Korea) by GS-FLX (Roche).…”

“…We have established a rock bream cDNA shotgun sequence transcriptome database using a next-generation sequencing (NGS) technology [23] and a BAC library was constructed as previously described [24]. Two distinct cDNAs which showed homology to known IkBa were identified from homology searches using the Basic Local Alignment Tool (BLAST) algorithm.…”

Molecular genomic- and transcriptional-aspects of a teleost TRAF6 homolog: Possible involvement in immune responses of Oplegnathus fasciatus against pathogens

“…The details of indoor animal rearing, multi-tissue collection and blood cell isolation followed by cDNA and BAC gDNA library construction were described previously [20,21]. The cDNA database search using BLAST tool yielded two partial contigs spanning the mid and 3'-proximal regions and exhibited significant similarity with known TRAF6 homologs.…”