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A Converging Strategy for the Generation of a Virtually Sequenced cDNA Library from Unreferenced Pacific Oysters
Published on: June 13, 2019
A tandem-repeat galectin involved in innate immune response of the pearl oyster Pinctada fucata
Zhongliang Wang1, Jichang Jian, Yishan Lu
1College of Fisheries, Guangdong Ocean University, Zhanjiang 524025, China.
Abstract:
The cDNA of a tandem-repeat galectin from the pearl oyster Pinctada fucata (designated PfGal) was cloned by expressed sequence tag (EST) and rapid amplification of cDNA ends (RACE) techniques. The full-length cDNA of PfGal was 1386bp, consisting of a 5' untranslated region (UTR) of 26bp, a 3' UTR of 313bp, and an open reading frame (ORF) of 1047bp encoding a polypeptide of 348 amino acids with a predicted molecular weight of 38.09kDa and theoretical isoelectric point of 8.49. Similar to other tandem-repeat galectins, PfGal contained two tandem carbohydrate recognition domains (CRDs), with typical conserved motifs which were important for carbohydrate recognition, and it appeared to possess neither a signal peptide nor a transmembrane domain. Fluorescent quantitative real-time PCR analyses indicated that PfGal mRNA was highly expressed in hemocytes, digestive gland and mantle, and its expression was increased in all studied tissues after Vibrio alginolyticus challenge. The temporal expression of PfGal mRNA in hemocytes challenged by V. alginolyticus was clearly time-dependent and reached the maximum level at 6h post-challenge, and then recovered to the original level. These results collectively indicated that PfGal may be involved in the immune response against bacterial infection and clearance of bacterial pathogens in P. fucata.
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