title: Molecular cloning, expression profiles and promoter analysis of insulin-like growth factor binding protein-4 (IGFBP-4) in Japanese flounder (Paralichthys olivaceus).
abstract: We cloned and characterized cDNA sequence of insulin-like growth factor binding protein-4 (IGFBP-4) from Japanese flounder (Paralichthys olivaceus). The 1493 bp full-length cDNA sequence contained an open reading frame (ORF) of 780 bp, which encoded a protein of 259 amino acids. The deduced amino acid sequences included a putative signal peptide of 28 amino acid residues resulting in a mature protein of 231 amino acids. Twenty cysteine residues and two conserved IGFBPs motif (GCGCCXXC and CWCV) were found in the N- and C-terminal domain. In the over 13 kbp genomic sequence, four exons, three introns, and 5'-/3'-flanking sequences were identified. Sequence alignment and phylogenetic analysis showed that Japanese flounder IGFBP-4 was indeed the ortholog of the human IGFBP-4 gene and shared high identities with other teleost IGFBP-4 genes. The promoter region was also analyzed and several potential transcription factor (TF) binding sites were determined which may modulate the IGFBP-4 expression. Quantitative real-time PCR analysis revealed that IGFBP-4 mRNA was observed in various tissues, with intestine showing the highest expression. The maternal transcripts of IGFBP-4 gene existed in the early embryonic stages and then increased in the following stages until hatching, suggesting that IGFBP-4 may be involved in the fish early development. The expression level of IGFBP-4 mRNA was relatively higher at 3 days post hatching (dph) and 15 dph, and gradually decreased during the metamorphosis period. All these results indicated that IGFBP-4 plays a significant role in IGF regulating vertebrate growth and development.
authors.affiliation: College of Marine Life Sciences, Ocean University of China, Key Laboratory of Marine Genetics and Breeding, Ministry of Education, #5 Yushan Road, Qingdao 266003, China.
authors.forename: Jing
authors.initials: J
authors.lastname: Wang
authors.affiliation: College of Marine Life Sciences, Ocean University of China, Key Laboratory of Marine Genetics and Breeding, Ministry of Education, #5 Yushan Road, Qingdao 266003, China.
authors.forename: Jinning
authors.initials: J
authors.lastname: Gao
authors.affiliation: College of Marine Life Sciences, Ocean University of China, Key Laboratory of Marine Genetics and Breeding, Ministry of Education, #5 Yushan Road, Qingdao 266003, China.
authors.forename: Wenji
authors.initials: W
authors.lastname: Wang
authors.affiliation: College of Marine Life Sciences, Ocean University of China, Key Laboratory of Marine Genetics and Breeding, Ministry of Education, #5 Yushan Road, Qingdao 266003, China.
authors.forename: Liman
authors.initials: L
authors.lastname: Ma
authors.affiliation: College of Marine Life Sciences, Ocean University of China, Key Laboratory of Marine Genetics and Breeding, Ministry of Education, #5 Yushan Road, Qingdao 266003, China.
authors.forename: Mengmeng
authors.initials: M
authors.lastname: Liu
authors.affiliation: College of Marine Life Sciences, Ocean University of China, Key Laboratory of Marine Genetics and Breeding, Ministry of Education, #5 Yushan Road, Qingdao 266003, China.
authors.forename: Haiyang
authors.initials: H
authors.lastname: Yu
authors.affiliation: College of Marine Life Sciences, Ocean University of China, Key Laboratory of Marine Genetics and Breeding, Ministry of Education, #5 Yushan Road, Qingdao 266003, China.
authors.forename: Zhigang
authors.initials: Z
authors.lastname: Wang
authors.affiliation: College of Marine Life Sciences, Ocean University of China, Key Laboratory of Marine Genetics and Breeding, Ministry of Education, #5 Yushan Road, Qingdao 266003, China.
authors.forename: Xubo
authors.initials: X
authors.lastname: Wang
authors.affiliation: College of Marine Life Sciences, Ocean University of China, Key Laboratory of Marine Genetics and Breeding, Ministry of Education, #5 Yushan Road, Qingdao 266003, China.
authors.forename: Jie
authors.initials: J
authors.lastname: Qi
authors.affiliation: College of Marine Life Sciences, Ocean University of China, Key Laboratory of Marine Genetics and Breeding, Ministry of Education, #5 Yushan Road, Qingdao 266003, China. Electronic address: qzhang@ouc.edu.cn.
authors.forename: Quanqi
authors.initials: Q
authors.lastname: Zhang
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D018076:DNA, Complementary / Q000235:genetics; D005432:Flounder / Q000196:embryology / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism; D018974:Insulin-Like Growth Factor Binding Protein 4 / Q000235:genetics* / Q000378:metabolism; D007814:Larva / Q000235:genetics / Q000254:growth & development / Q000378:metabolism; D008969:Molecular Sequence Data; D009928:Organ Specificity; D010802:Phylogeny; D011401:Promoter Regions, Genetic
chemical_list: D018076:DNA, Complementary; D018974:Insulin-Like Growth Factor Binding Protein 4
keywords: Insulin-like growth factor binding protein-4; Paralichthys olivaceus; Promoter analysis; cDNA cloning; mRNA expression