Molecular cloning and tissue and temporal expression of PTEN gene in early development of goldstriped amberjack Seriola aureovittata
SUN Ran-ran1, SHI Bao2, LIU Xue-zhou1,2, CHANG Ya-qing1, ZHANG Yan-xiang3, GAO Quan-yi3, XU Yong-jiang2, WANG Bin2, JIANG Yan2, ZHANG Zheng-rong2
1.College of Fisheries and Life Science, Dalian Ocean University, Dalian 116023, China; 2.Qingdao National Laboratory for Marine Science and Technology,Laboratory for Marine Fisheries Science and Food Production Processes, Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao 266071, China; 3.Dalian Fugu Fishery Company Limited, Dalian 116400, China
Abstract: The full-length cDNA sequence of the growth regulator PTEN was cloned by homologous cloning and rapid amplification of cDNA ends (RACE),and temporal and spatial expression of PTEN mRNA was analyzed in goldstriped amberjack Seriolaaureovittata by quantitative PCR (qPCR) to explore the molecular mechanism of PTEN in the growth and development of goldstriped amberjack.The results showed that the PTEN cDNA sequence was 1575 bp in length and the ORF was 1287 bp, encoding 428 amino acids.PTEN has the common feature of vertebrate PTEN and contains PTP (PTPc motif) and PTEN-C2 domain.Sequence alignment analysis revealed that both goldstriped amberjack and greater amberjack Serioladumerili had a high homology(95%).The phylogenetic tree indicated that goldstriped amberjack PTEN was clustered into one branch with the PTEN Perciformes and Pleuronectiformes.By qPCR analysis, the maximal relative expression of PTEN was observed in the heart of goldstriped amberjack (P<0.05), followed by in the spleen, head kidney, kidney, stomach, intestine, gill and pituitary, indicating that the PTEN is involved in the regulation of a variety of physiological processes in various tissues due to high expression of PTEN mRNA.It was found that PTEN was continuously expressed in early embryonic development(embryonic stage, larvae and juvenile), with significantly higher expression level from early blastula stage (38 hours post fertilization, hpf) (P<0.05) with the maximal expression level in freshly hatched larvae (P<0.05).The expression level of PTEN mRNA was gradually decreased during the postlarvae and juvenile (P<0.05).The expression level of PTEN was found to be gradually decreased, significantly and constantly low at 35 days old.The findings indicate that PTEN plays an important regulatory role in the development of embryos and juveniles, which provides a theoretical basis with research of the regulation mechanism and culture during the early growth and development stages in goldstriped amberjack.