Gene encoding prolactin of red-spotted grouper, Epinephelus akaara, and its application as a molecular marker for grouper species identification
1
조회
0
다운로드
0
인용
0
참고문헌
키워드
초록
Groupers are economically important species in the fishery and aquaculture industries in Asian countries. Various species of grouper, including hybrids, have been brought to market even without precise species identification. In this study, we analyzed the structure and expression profile of the gene encoding prolactin (PRL) in the red-spotted grouper Epinephelus akaara based on genomic DNA and cDNA templates. The results showed that the PRL gene consists of five exons encoding an open reading frame of 212 amino acids, including a putative signal peptide of 24 amino acids and a mature structural protein of 188 amino acids. It showed amino acid identities of 99% with Epinephelus coioides, 83% with Amphiprion melanopus, 82% with Acanthopagrus schlegelii, 75% with Oreochromis niloticus, 70% with Coregonus autumnalis, and 67% with Oncorhynchus mykiss, indicating its closer similarity to E. coioides and other groupers but marked distinction from non-teleost PRLs. PRL mRNA expression was detected mostly in the brain, including the pituitary gland, with little expression in other tissues. While the 5-exon structure of the PRL gene of red-spotted grouper and the exon sizes were conserved, the sizes of the introns, particularly the first intron, were markedly different among the grouper species. To examine whether these differences can be used to distinguish groupers of similar phenotypes, exon-primed intron-crossing analysis was carried out for various commercially important grouper species. The results showed clear differences in size of the amplified fragment encompassing the first intron of the PRL gene, indicating that this method could be used to develop species-specific markers capable of discriminating between grouper species and their hybrids at the molecular level.
Abstract
Groupers are economically important species in the fishery and aquaculture industries in Asian countries. Various species of grouper, including hybrids, have been brought to market even without precise species identification. In this study, we analyzed the structure and expression profile of the gene encoding prolactin (PRL) in the red-spotted grouper Epinephelus akaara based on genomic DNA and cDNA templates. The results showed that the PRL gene consists of five exons encoding an open reading frame of 212 amino acids, including a putative signal peptide of 24 amino acids and a mature structural protein of 188 amino acids. It showed amino acid identities of 99% with Epinephelus coioides, 83% with Amphiprion melanopus, 82% with Acanthopagrus schlegelii, 75% with Oreochromis niloticus, 70% with Coregonus autumnalis, and 67% with Oncorhynchus mykiss, indicating its closer similarity to E. coioides and other groupers but marked distinction from non-teleost PRLs. PRL mRNA expression was detected mostly in the brain, including the pituitary gland, with little expression in other tissues. While the 5-exon structure of the PRL gene of red-spotted grouper and the exon sizes were conserved, the sizes of the introns, particularly the first intron, were markedly different among the grouper species. To examine whether these differences can be used to distinguish groupers of similar phenotypes, exon-primed intron-crossing analysis was carried out for various commercially important grouper species. The results showed clear differences in size of the amplified fragment encompassing the first intron of the PRL gene, indicating that this method could be used to develop species-specific markers capable of discriminating between grouper species and their hybrids at the molecular level.
참고문헌 (34)
- Astola A, "Isolation ofSparus auratus prolactin gene and activity of the cis-acting regulatory elements", Gen Comp Endocrinol, 134, pp.57~61 (2003)
- Breves JP, "Osmoregulatory actions of prolactin in the gastrointestinal tract of fishes", General and Comparative Endocrinology, pp.113589~ (2020)
- Chang YS, "Molecular cloning of silver carp and bighead carp prolactin", Gen Comp Endocrinol, 87, pp.260~265 (1992)
- Chang YJ, "Black porgy(Acanthopagrus schlegeli)prolactin cDNA sequence : mRNA expression and blood physiological responses during freshwater acclimation", Comp Biochem Physiol B Biochem Mol Biol, 147, pp.122~128 (2007)
- Chao SC, "Nucleotide sequence of carp prolactin cDNA", Nucleic Acids Res, 16, pp.9350~ (1988)
- Cooke NE, "Human prolactin : cDNA structural analysis and evolutionary comparisons", J Biol Chem, 256, pp.4006~4016 (1981)
- Eschmeyer WN, "Genera in the family Serranidae. Catalog of Fishes" (2010), California Academy of Sciences
- Gong N, "Discovery of prolactin-like in lamprey : role in osmoregulation and new insight into the evolution of the growth hormone", prolactin family, 2022, pp.119, e2212196119~
- Hebert PDN, "Biological identifications through DNA barcodes", Proc R Soc Lond B, 270, pp.313~321 (2003)
- Inoue H, "High efficiency transformation ofEscherichia coli with plasmids", Gene, 96, pp.23~28 (1990)
- Kang MJ, "The embryonic development and hatchability of two hybrids with giant grouper female : giant grouper ♀×kelp grouper ♂ and giant grouper ♀×red-spotted grouper ♂", Korean J Fish Aquat Sci, 53, pp.557~562 (2020)
- Lee CH, "Induced maturation and fertilized egg production of the red-spotted grouper, Epinephelus akaara, using adaptive physiology of photoperiod and water temperature", Aquac Res, 51, pp.2084~2090 (2020)
- Lee KM, "Prolactin and prolactin receptor expressions in a marine teleost, pufferfishTakifugu rubripes", Gen Comp Endocrinol, 146, pp.318~328 (2006)
- Li C, "Exon-primed intron-crossing(EPIC)markers for non-model teleost fishes", Evol Biol, 10, pp.90~ (2010)
- Lim SG, "Effects of salinity on the growth, survival and stress responses of red spotted grouperEpinesphelus akaara and hybrid grouperE. akaara ♀ ×E. lanceolatus ♂", Korean J Fish Aquat Sci, 49, pp.612~619 (2016)
- Mai W, "Cloning and immune characterization of the c-type lysozyme gene in red-spotted grouper, Epinephelus akaara", Fish Shellfish Immunol, 36, pp.305~314 (2014)
- Manzon LA, "The role of prolactin in fish osmoregulation : a review", Gen Comp Endocrinol, 125, pp.291~310 (2002)
- Monaghan MT, "DNA-based species delineation in tropical beetles using mitochondrial and nuclear markers", Philos Trans R Soc Lond B Biol Sci, 360, pp.1925~1933 (2005)
- Noh CH, "Hatchability of fertilized eggs from grouper(subfamily Epinephelinae)hybrids in Korea : a mini review for selection of commercially promising cross combinations", Korean J Fish Aquat Sci, 53, pp.479~485 (2020)
- Noh GE, "Characterization of genes encoding prolactin and prolactin receptors in starry flounderPlatichthys stellatus and their expression upon acclimation to freshwater", Fish Physiol Biochem, 39, pp.263~275 (2013)
- Noh GE, "Gene encoding prolactin in cinnamon clownfishAmphiprion melanopus and its expression upon acclimation to low salinities", Aquat Biosyst, 9, pp.1~ (2013)
- Park YC, "Development of detection method forNiphon spinosus, Epinephelus bruneus, andEpinephelus septemfasciatus using 16S rRNA gene", Korean J Food Sci Technol, 45, pp.1~7 (2013)
- Quérat B, "Sequence and regulation of European eel prolactin mRNA", Mol Cell Endocrinol, 102, pp.151~160 (1994)
- Rentier-Delrue F, "Tilapia prolactin : molecular cloning of two cDNAs and expression inEscherichia coli", DNA, 8, pp.261~270 (1989)
- Rimmer MA, "A review of grouper(family Serranidae : subfamily Epinephelinae)aquaculture from a sustainability science perspective", Rev Aquac, 11, pp.58~87 (2019)
- Sadovy Y, "Epinephelus akaara The IUCN red list of threatened species" (2018), International Union for Conservation of Nature(IUCN)
- Saitou N, "The neighbor-joining method : a new method for reconstructing phylogenetic trees", Mol Biol Evol, 4, pp.406~425 (1987)
- Si Y, "Isolation of prolactin gene and its differential expression during metamorphosis involving eye migration of Japanese flounderParalichthys olivaceus", Gene, 780, pp.145522~ (2021)
- Song S, "Molecular cloning and expression of salmon prolactin cDNA", Eur J Biochem, 172, pp.279~285 (1988)
- Swennen D, "Structure of the tilapia(Oreochromis mossambicus)prolactin I gene", DNA Cell Biol, 11, pp.673~684 (1992)
- Varma S, "Cloning and nucleotide sequence of ovine prolactin cDNA", Gene, 77, pp.349~359 (1989)
- Xiong F, "A gene encoding chinook salmon(Oncorhynchus tschawytscha)prolactin : gene structure and potential cis-acting regulatory elements", Mol Mar Biol Biotechnol, 1, pp.155~164 (1992)
- Yamaguchi K, "Complete amino acid sequences of a pair of fish(tilapia)prolactins, tPRL177 and tPRL188", J Biol Chem, 263, pp.9113~9121 (1988)
- Zhang W, "cDNA sequence and spatio-temporal expression of prolactin in the orange-spotted grouper, Epinephelus coioides", Gen Comp Endocrinol, 136, pp.134~142 (2004)