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Liu H.,Shantou University | Liu H.,Mariculture Research Center for Subtropical Shellfish & Algae | Guo Z.,Shantou University | Guo Z.,Mariculture Research Center for Subtropical Shellfish & Algae | And 9 more authors.
Molecular Biology Reports | Year: 2014

Long-chain polyunsaturated fatty acids (LC-PUFAs) are essential in lots of important physiological processes, while, many marine species have no or limited ability of endogenous PUFA biosynthesis, normally due to the lack of key enzymes such as fatty acid desaturase (FAD). In this study, we isolated a scallop Chlamys nobilis cDNA with high homology to vertebrate FADs. Functional characterization in recombinant yeast Saccharomyces cerevisiae showed that scallop FAD exhibited Δ5-desaturation activity towards both saturated and PUFA substrates. Thus, it efficiently desaturated exogenously added PUFA C20:4(n − 3) and C20:3(n − 6) to C20:5(n − 3) (EPA) and C20:4(n − 6) (ARA) respectively. It also converted the yeast’s endogenous C18:0 into C18:1(n − 13), and participated in the biosynthesis of non-methylene-interrupted FA by introducing a double bond to C20:3(n − 3) and C20:2(n − 6) in the Δ5 carbon. Temporal transcript profile of scallop FAD was studied during early embryonic development. High level of mRNA was found at the beginning of embryogenesis (egg) and noticeable decreases of were observed during larvae development, suggesting maternal FAD mRNA transfer to the embryo. Further, FAD transcripts were detected in all tissues analyzed, with the gonad and hepatopancreas showing the highest expression. © 2014, Springer Science+Business Media Dordrecht. Source

Wang Y.-J.,Shantou University | Wang Y.-J.,Mariculture Research Center for Subtropical Shellfish & Algae | Zheng H.-P.,Shantou University | Zheng H.-P.,Mariculture Research Center for Subtropical Shellfish & Algae | And 8 more authors.
Molecular Biology Reports | Year: 2014

Cold shock domain proteins (CSDPs) play various roles in cellular processes, including adaptation to low temperature in mollusk. In this study, the cDNA encoding CSDP in noble scallop Chlamys nobilis was cloned (CnCSDP), and its transcript profile in cold-stressed orange and brown strains was analyzed using real-time RT-PCR. Bioinformatic analysis shows that the complete CnCSDP cDNA is 1,434 bp in length, consisting of a 5′-, and 3′-untranslated region of 86 and 466 bp, respectively, and a 882 bp open reading frame encoding 294 amino acids with an estimated molecular mass of 33.23 kDa and an isoelectric point of 9.72. Furthermore, CnCSDP has two consensus RNA binding domains, four DNA binding sites and four RGG repeats. CnCSDP transcripts amount in the blood of orange strain were significantly higher, while in the mantle were much lower than that of brown strain. In the cold-stressed orange strain, CnCSDP mRNA decreased dramatically in the blood and mantle, but was significantly up-regulated at 1 h, and progressively returned to its original levels in the gill. In the cold-stressed brown strain, CnCSDP transcripts were significantly up-regulated at 6 h in the blood, decreased dramatically in the mantle, while mostly maintained unchanged in the gill. Lastly, significantly positive correlation was found between the CnCSDP transcripts amount and total carotenoids content in the gill in both orange and brown strain. This study will be helpful for understanding the molecular mechanism underlying cold stress in noble scallop in the further. © 2014, Springer Science+Business Media Dordrecht. Source

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