Bin LIU, Xiao-Nan ZANG, Shun-Mei LIU, Xue-Cheng ZHANG, Ji-Lin LEI

Cloning, sequence characterization and phylogenetic analysis on full- lengthcdna of growth hormone from southern flounder (Paralichthys lethostigma)

Pisibalığının(Paralichthyslethostigma) DNA’sından sentezlenen (SMART) RACE RNA’nın 5’ ucunda büyüme hormonundan (PLGH) kodlanan cDNA, anahtar mekanizması ile klonlanmıştır. Sekans, 190 amino asitli polipeptid kodlayan 902 nükleotidli, 17 amino asitli tek bir peptidtir. Messenger RNA’nın dönüşmeyen bölgesinin (UTR; untranslatedregion) 5’ ve 3’ uçları sırasıyla 131 ve 198 nükleotidlidir. Çıkarılabilir amino asit sekanslarının diğer Pleuronectiformes türleri ile birlikte sıralanmasından sonra, Paralichthyidae’nın üç türünün tümünde de C ile sonlanan ardışık 14 amino asit segmentinin olmadığı bulundu. Pleuronectiformes türleri arasında Paralichthyidae’nın büyüme hormonunun (GH) en kısa boya sahip olduğu kanıtlanabilir. Benzer analizler, PLGH’nincDNA’sınınParalichthysorbignyanu’da en yüksek skora, Soleasenegalensis’de en düşük skora sahip olduğunu göstermiştir. Büyüme hormonunun ORF (açık okuma çerçevesi) sekansının tamamında filogenetik analizler maksimum sadeleştirilmiş metot kullanılarak yapılmıştır. Topolojide türler üç gruba ayrılır: Soleoidei,Paralichthyidae ve Pleuronectidae. Bu çalışmada Soleoidei’nin en seçkin türü seçilmiş, Soleasenegalensis, bağımsız bir sınıf oluşturmuş ve Paralichthyidae ve Pleuronectidae’dan nispeten yüksek bir ortalama fark göstermiştir. Büyüme hormonunun (GH) ORF sekansına dayalı filogenetik analizlerin sonuçları, Pleuronectiformesfilogenide büyüme hormonunun (GH) ORF sekansındaki güvenilir potansiyel uygulamaları öneren Pleuronectiformes türlerinin 12S, 16S rDNA ve mitokondrial DNA sekanslarının daha önceki sonuçları ile örtüşmektedir.

Anahtar Kelimeler:

Paralichthys, polipeptidler, somatotropin, balık, transkripsiyon, amino asit dizileri, tamamlayıcı DNA, DNA klonlama, elçi RNA, nükleotit dizileri, filogeni

Güney pisibalığının (Paralichthys lethostigma) büyüme hormonundaki cdna (tamamlayıcı dna) üzerinde klonlama, sekans tanımlanmasıve filogenetik analizler

The full-length cDNA encoding growth hormone (PLGH) of Southern flounder (Paralichthyslethostigma) was cloned by Switching Mechanism At 5' end of the RNA Transcript (SMART) RACE. The sequence was 902 nucleotides long, coding a polypeptide of 190 amino acids and a signal peptide of 17 amino acids. The 5’ and 3’ UTR of the messenger RNA were 131 and 198 nucleotides long respectively. After alignment of the deduced amino acid sequence with other Pleuronectiformes species, a consecutive absence of 14 amino acids segment in the C-terminal was found in all three species ofParalichthyidae. It can be proved that GHs of Paralichthyidae have the shortest length in Pleuronectiformes species. Homology analysis of PLGH cDNA showed the highest score with Paralichthysorbignyanuand the lowest score with Soleasenegalensis. The phylogenetic analysis on complete GH ORF sequences was performed with the maximum parsimony method. In the topology founded, the species fell into three groups: Soleoidei,Paralichthyidae and Pleuronectidae. The exclusive species ofSoleoidei selected in this work,Soleasenegalensis formed an independent branch and showed relatively high mean difference with Paralichthyidae and Pleuronectidae. The results of phylogenetic analysis based on the GH ORF sequences were consistent with the earlier results based on 12S, 16S rDNA and mitochondrial DNA sequences within Pleuronectiformes species, which would suggest the potential credible application of GH ORF sequences in Pleuronectiformes phylogeny.

Keywords:

Paralichthys, polypeptides, somatotropin, fishes, transcription, amino acid sequences, complementary DNA, DNA cloning, messenger RNA, nucleotide sequences, phylogeny,

PDF

___

Acosta, J., Carpio, Y., Besada, V., Morales, R., Sanchez, A., Curbelo, Y., Ayala, J and Estrada, M.P. 2008. Recombinant truncated tilapia growth hormone enhances growth and innate immunity in tilapia fry (Oreochromissp), General and Comparative Endocrinology, 157: 49-57. doi:10.1016/j.ygcen.2008.03.009
Agelon, L.B. 1998. Promotion of rapid growth of rainbow trout (Salmo gairdneri) by a recombinant fish growth hormone.Canadian Journal of Fisheries and Aquatic Sciences, 45: 146-151. doi: 10.1139/f88-016
Almuly, R., Cavari, B., Ferstman, H., Kolodny, O. andFunkenstein, B. 2000. Genomic structure and sequence of the gilthead seabream (Sparusaurata) growth hormone-encoding gene: identification of minisatellite polymorphism in intronI.Genome, 43: 836-845.
Benetti, D.D. 2000. Aquaculture of Southern flounder, Paralichthyslethostigma, in freshwater recirculating systems.The Advocate, Global Aquaculture Alliance Technical Magazine, 3(2): 20.
Berendzen, P.B. andDimmick, W.W. 2002. Phylogenetic relationships of Pleuronectiformes based on molecular evidence. Copeia, 3:642–652.
Bernardi, G.D., Onofrio, G. andCaccio, S. 1993. Molecular phylogeny of bony fishes, based on the amino acid sequence of the growth hormone.Journal of Molecular Evolution, 37: 644-649. doi: 10.1007/BF00182750
Byrappa,V. and Sydney, B. 1997. Genomic structure and sequence of the pufferfish (Fugurubripes) growth hormone-encoding gene: a comparative analysis of teleost growth hormone genes. Gene, 187: 211-215.
Carlos, I., Gaetano, C. and Manuel, M. 2004. Phylogenetic Relationships Among Ten Sole Species (Soleidae, Pleuronectiformes) from the Gulf of Ca´diz (Spain) Based on Mitochondrial DNA Sequences. Marine Biotechnology, 6: 612–624.
Cavari, B., Funkenstein, B.and Chen, T.T. 1993. Effect of growthhormone on the growth rate of the gilthead seabream (Sparusaurata) and the use of different constructs for production of transgenic fish. Aquaculture, 111: 189-197. doi:10.1016/0044-8486(93)90036-X
Chen, Y., Wang, Y.P., He, S.P. and Zhu, Z.Y. 2004. Cloning and Sequencing of the Growth Hormone Gene of Large Yellow Croaker and Its Phylogenetic Significance. Biochemical Genetics, 42: 9-10.
Clements, M.D., Bart Jr, H.L. and Hurley, D.L. 2004.Isolation and characterization of two distinct growth hormonecDNAs from the tetraploid smallmouth buffalofish (Ictiobusbubalus).General and Comparative Endocrinology, 136: 411-418.
De Jesus, E.G.T.,Ayson, F.G., Amemiya, Y.,Moriyama, S., Hyodo, S., Hirano, T. andKawauchi, H. 2002.Milkfish (Chanoschanos) growth hormone cDNAcloning and mRNA expression in embryos andearly larval stages, Aquaculture, 208: 177-188. doi:10.1016/S0044-8486(01)00759-1
Fausto, T., Andrea, C., Maria, V., Corrado, P. and Anna Maria, S. 1999. Comparative Analysis of a Mitochondrial DNA Control Region Fragment Amplified from Three Adriatic Flatfish Species and Molecular Phylogenesis of Pleuronectiformes.Marine Biotechnology, 1: 20-24.
Favre-Krey, L., Theodoridou, M., Boukouvala E., Panagiotidis, C.H.,Papadopoulos, A.I.,Sklaviadis, T. andKrey, G. 2007. Molecular characterization of a cDNA from the gilthead sea bream(Sparusaurata) encoding a fish prion protein.Comparative Biochemistry and Physiology Part B: Biochemistry and Molecular Biology, 147(3): 566-573. doi:10.1016/j.cbpb.2007.03.011
Forsyth, I.A. and Wallis, M. 2002. Growth hormone and prolactin: molecular and functional evolution. Journal of Mammary Gland Biology and Neoplasia, 7: 291-312.
Infante, C., Catanese, G. andManchado, M. 2004. Phylogenetic relationships among ten species of soles (Soleidae,Pleuronectiformes) from the Gulf of Cádiz (Spain) based on mitochondrial DNA sequences. Marine Biotechnology, 6: 612–624.
Li, Y.H., Bai, J.J., Jian, Q., Ye, X., Lao, H.H., Li, X.H., Luo, J.R., Liang, X.F. 2003. Expression of common carp growth hormone in the yeast Pichiapastorisand growth stimulation of juvenile tilapia (Oreochromisniloticus).Aquaculture, 216: 329-341. doi:10.1016/S0044-8486(02)00406-4
Marins, L.F., Levy, J.A., Folch, J.M and Sanchez, A.2003.A growth hormone-based phylogenetic analysis of euteleostean fishesincluding a representative species of the Atheriniformes Order,Odontesthesargentinensis, Genetics and Molecular Biology, 26(3): 295-300.doi: 10.1590/S1415-47572003000300013
McLean, E.D., Teskeredzic, E. and Souga, L.M. 1993. Growth enhancement following ietary delivery recombinant protein somatotropin to diploid and triploid coho salmon (Oncorhynchus kisutch).FishPhysiology and Biochemistry, 11: 363-369. doi: 10.1007/BF00004586
Pardo, B.G., Machordom, A., Foresti, F., Porto-Foresti, F., Azevedo, M.F.C., Banon, R., Sanchez, L. and Martinez, P. 2005. Phylogenetic analysis of flatfish (Order:Pleuronectiformes) based on mitochondrial 16s rDNA sequences. Scientia Marina, 69:531–543.
Pendon, C., Martinez-Barbera, J.P., Perez-Sanchez, J., Roudriguez, R.B., Grenett, H. and Valdivia, M.M. 1994. Cloning of sole (Soleasenegalensis) growth hormone-encoding cDNA.Gene, 145: 273–240.
Peyush, P., Shunsuke, M., Akiyoshi, T. and Hiroshi, K. 2000. Molecular Cloning of Growth Hormone Complementary DNA in Barfin Flounder (Veraspermoseri). Marine Biotechnology, 2: 21–26. doi: 10.1007/s101269900004
Rubin, D.A. andDores, R.M. 1994. Cloning of a growth hormone from a primitive bony fish and its phylogenetic relationships.General and Comparative Endocrinology, 95: 71-83. doi:10.1006/gcen.1994.1103
Rubin, D.A. andDores, R.M. 1995. Obtaining a more resolute teleost growth hormone phylogeny by the introduction of gaps in sequence alignment.Molecular Phylogenetics and Evolution, 4: 129-138.
Rubin, D.A., Youson, J.H., Marra, L.E. andDores, R.M. 1996. Cloning of a gar (Lepisosteusosseus) GH cDNA: trends in actinopterygian GH structure.Journal of Molecular Endocrinology, 16: 73-80. doi: 10.1677/jme.0.0160073
Somers, W., Ultsch, M., de Vos, A.M. andKossiakof, A.A. 1994.The Xray structure of a growth hormone-prolactin receptor complex.Nature, 372(6505): 478-481. doi: 10.1038/372478a0
Suzuki, T., Srivastava, A.S. andKurokawa, T. 2002.cDNA cloning and phylogenetic analysis of pancreatic serine proteases from Japanese flounder, Paralichthysolivaceus. Comparative Biochemistry and Physiology Part B: Biochemistry and Molecular Biology, 131: 63–70. doi:10.1016/S1096-4959(01)00487-0
Swofford, D.L. 2000.PAUP (Phylogenetic analysis using parsimony)and other methods.Version 4.Sinauer Associates, Sunderland, Massachusetts.
Tanaka, M., Hosokawa, Y., Watahiki, M., Nakashima, K., 1992.Structure of the chicken growth hormone-encoding gene and its promoter region. Gene 112, 235–239.doi:10.1016/0378-1119(92)90382-Y
Tsai, H.J., Kuo, J.C., Lou, S.W. and Kuo, T.T. 1994. Growth enhancement of juvenile striped mullet by feeding recombinant yeast containing fish growth hormone.Progressive Fish Culturist, 56: 7-12.
Watahiki, M., Yamamoto, M., Yamakawa, M., Tanaka, M. and Nakashima, K. 1989. Conserved and unique amino acid residues in the domains of the growth hormones. Flounder growth hormone deduced from the cDNA sequence has the minimal size in the growth hormone prolactin gene family.Journal of Biological Chemistry, 264: 312- 316.
Watahiki, M., Ohara, H., Tsuda, M., Shouji, K., Masuji, A., Tanaka, M., Yamakawa, M., Ushiro, H., Yoneda, Y. and Nakashima, K. 1992.Synthesis of recombinant yellowtail and flounder growth hormone in Escherichia coli.Bioscience, Biotechnology and Biochemistry, 56: 1012-1016.
Wenner, C.A., Roumillat, W.A., Moran, J.E., Maddox, M.B., Daniel III, L.B and Smith, J.W. 1990.Investigations on the life history and population dynamics of marine recreational fishes in South Carolina: Part 1. Marine Resources Research Institute, South Carolina Wildlife and Marine Resources Department, Charleston, SC. 177 pp.
Zang, X.N., Liu, B., Liu, S.M., Sun, P.N., Zhang, X.Q. and Zhang, X.C. 2007.Transformation and expression of Paralichthysolivaceus growth hormone cDNA in Synechocystis sp. PCC6803.Aquaculture, 266: 63-69. doi:10.1016/j.aquaculture.2007.02.027
Zhang, J.N., Song, P., Hu, J.R., Mo, S.J., Peng, M.Y., Zhou, W., Zou, J.X. and Hu, Y.C. 2005. Molecular Cloning and Sequence Analysis of Full-Length Growth Hormone cDNAs from Six Important Economic Fishes, ActaGeneticaSinica, 32(1): 19-29.