title: Hsp60 in caudal fin regeneration from Paramisgurnus dabryanus: molecular cloning and expression characterization.
abstract: Heat shock protein 60 (Hsp60) is a kind of highly conserved immunogenic molecule involved in a wide range of biochemical processes in response to external stressors. Its multifunction in regulating immune responses and modulating signal pathway interests us in investigating its role in fin regeneration that has become an excellent and interesting model for studying the molecular basis of morphogenesis. We firstly clarified basical process and crucial period of caudal fins regeneration in Paramisgurnus dabryanus by histological analysis. Then we cloned full-length cDNA of hsp60 from P. dabryanus (designated as PdHsp60) by RACE method. The cDNA contains a 124 bp 5'UTR, a 1731 bp open reading frame (ORF) encoding 576 amino acids and a 510 bp 3'UTR (Accession no.: KF544774). The phylogenetic tree shows that the PdHsp60 fits within the hsp60 clade. And quantitative RT-PCR detected the PdHsp60 began to increase rapidly its expression at 1 dpa and reached its peak at 2 dpa. Next, spatial distribution analysis of PdHsp60 in fins showed that PdHsp60 located mainly in the deeper lay of regenerated epidermis when PdHsp60 expressed most. After the PdHsp60 had been cloned into the pET-32a vector, SDS-PAGE and Western blotting analysis confirmed that the PdHsp60 protein was efficiently expressed in Escherichia coli BL21. These findings have revealed that PdHsp60, a highly conserved gene related to the innate immune system and stress response during vertebrate evolution, is involved in response to wounding stimulation--in the formation of wound epidermis which occurs as the first phase of fin regeneration after fin amputation in caudal fin regeneration.
authors.affiliation: Molecular and Genetic Laboratory, College of Life Science, Henan Normal University, 46# East of Construction Road, Xinxiang, Henan 453007, China.
authors.forename: Li
authors.initials: L
authors.lastname: Li
authors.affiliation: Molecular and Genetic Laboratory, College of Life Science, Henan Normal University, 46# East of Construction Road, Xinxiang, Henan 453007, China.
authors.forename: Shengna
authors.initials: S
authors.lastname: Zhai
authors.affiliation: Molecular and Genetic Laboratory, College of Life Science, Henan Normal University, 46# East of Construction Road, Xinxiang, Henan 453007, China.
authors.forename: Lele
authors.initials: L
authors.lastname: Wang
authors.affiliation: Molecular and Genetic Laboratory, College of Life Science, Henan Normal University, 46# East of Construction Road, Xinxiang, Henan 453007, China.
authors.forename: Songbo
authors.initials: S
authors.lastname: Si
authors.affiliation: Molecular and Genetic Laboratory, College of Life Science, Henan Normal University, 46# East of Construction Road, Xinxiang, Henan 453007, China.
authors.forename: Hailan
authors.initials: H
authors.lastname: Wu
authors.affiliation: Molecular and Genetic Laboratory, College of Life Science, Henan Normal University, 46# East of Construction Road, Xinxiang, Henan 453007, China. Electronic address: changzhongjie@tom.com.
authors.forename: Zhongjie
authors.initials: Z
authors.lastname: Chang
mesh_terms: D000595:Amino Acid Sequence; D058500:Animal Fins / Q000502:physiology*; D000818:Animals; D001483:Base Sequence; D018834:Chaperonin 60 / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D003001:Cloning, Molecular; D003531:Cypriniformes / Q000145:classification / Q000235:genetics / Q000502:physiology*; D018076:DNA, Complementary / Q000235:genetics / Q000378:metabolism; D029941:Fish Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation; D008969:Molecular Sequence Data; D010802:Phylogeny; D012313:RNA / Q000235:genetics / Q000378:metabolism; D012038:Regeneration; D016415:Sequence Alignment / Q000662:veterinary
chemical_list: D018834:Chaperonin 60; D018076:DNA, Complementary; D029941:Fish Proteins; D012313:RNA
keywords: Fin regeneration; Gene expression; Paramisgurnus dabryanus; Wounding stimulation; hsp60 gene