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完整後設資料紀錄
DC 欄位 | 值 | 語言 |
---|---|---|
dc.contributor.advisor | 蕭仁傑(Jen-Chieh Hsiao) | |
dc.contributor.author | Po-Shun Chuang | en |
dc.contributor.author | 莊博舜 | zh_TW |
dc.date.accessioned | 2021-06-16T08:20:46Z | - |
dc.date.available | 2014-03-08 | |
dc.date.copyright | 2014-03-08 | |
dc.date.issued | 2014 | |
dc.date.submitted | 2014-01-29 | |
dc.identifier.citation | ANDRICH, F., CARNIELLI, J. B., CASSOLI, J. S., LAUTNER, R. Q., SANTOS, R. A., PIMENTA, A. M., DE LIMA, M. E. & FIGUEIREDO, S. G. 2010. A potent vasoactive cytolysin isolated from Scorpaena plumieri scorpionfish venom. Toxicon, 56, 487-96.
AUERBACH, P. S. 1991. Marine Envenomations. New England Journal of Medicine, 325, 486-493. BARON, A., DIOCHOT, S., SALINAS, M., DEVAL, E., NO L, J. & LINGUEGLIA, E. 2013. Venom toxins in the exploration of molecular, physiological and pathophysiological functions of acid-sensing ion channels. Toxicon, 75, 187-204. BLONDELLE, S. E. & HOUGHTEN, R. A. 1991. Hemolytic and antimicrobial activities of the twenty-four individual omission analogues of melittin. Biochemistry, 30, 4671-8. BURNETTE, W. N. 1981. 'Western Blotting' - Electrophoretic Transfer of Proteins from Sodium Dodecyl Sulfate-Polyacrylamide Gels to Unmodified Nitrocellulose and Radiographic Detection with Antibody and Radioiodinated Protein-A. Analytical Biochemistry, 112, 195-203. CAMERON, A. M. & ENDEAN, R. 1972. The venom glands of teleost fishes. Toxicon, 10, 301-3. CAMERON, A. M. & ENDEAN, R. 1973. Epidermal secretions and the evolution of venom glands in fishes. Toxicon, 11, 401-10. CARRIJO, L. C., ANDRICH, F., DE LIMA, M. E., CORDEIRO, M. N., RICHARDSON, M. & FIGUEIREDO, S. G. 2005. Biological properties of the venom from the scorpionfish (Scorpaena plumieri) and purification of a gelatinolytic protease. Toxicon, 45, 843-50. CHEN, D. S., KINI, R. M., YUEN, R. & KHOO, H. E. 1997. Haemolytic activity of stonustoxin from stonefish (Synanceja horrida) venom: pore formation and the role of cationic amino acid residues. Biochemical Journal, 325, 685-691. CHEN, W. J., BONILLO, C. & LECOINTRE, G. 2003. Repeatability of clades as a criterion of reliability: a case study for molecular phylogeny of Acanthomorpha (Teleostei) with larger number of taxa. Molecular Phylogenetics and Evolution, 26, 262-288. CHOMCZYNSKI, P. 1993. A Reagent for the Single-Step Simultaneous Isolation of RNA, DNA and Proteins from Cell and Tissue Samples. Biotechniques, 15, 532-&. CHOMCZYNSKI, P. & SACCHI, N. 1987. Single-Step Method of RNA Isolation by Acid Guanidinium Thiocyanate Phenol Chloroform Extraction. Analytical Biochemistry, 162, 156-159. CHURCH, J. E. & HODGSON, W. C. 2002. The pharmacological activity of fish venoms. Toxicon, 40, 1083-93. CUSHING, D. H. 1984. Fishes of the World, 2nd Edition - Nelson,Js. Nature, 312, 679-679. CUVIER, G. & LATREILLE, P. A. 1829. Le regne animal distribue d'apres son organisation: Les crustaces, arachnides et partie des insectes, par M. Latreille, Deterville. DUTERTRE, S. & LEWIS, R. J. 2010. Use of Venom Peptides to Probe Ion Channel Structure and Function. Journal of Biological Chemistry, 285, 13315-13320. ENDEAN, R. 1961. A Study of Distribution, Habitat, Behaviour, Venom Apparatus, and Venom of the Stone-Fish. Marine and Freshwater Research, 12, 177-190. FAY, J. C., WYCKOFF, G. J. & WU, C. I. 2001. Positive and negative selection on the human genome. Genetics, 158, 1227-1234. FELSENSTEIN, J. 1981. Evolutionary trees from DNA sequences: a maximum likelihood approach. Journal of molecular evolution, 17, 368-376. GARNIER, P., DUCANCEL, F., OGAWA, T., BOULAIN, J. C., GOUDEY-PERRIERE, F., PERRIERE, C. & MENEZ, A. 1997a. Complete amino-acid sequence of the beta-subunit of VTX from venom of the stonefish (Synanceia verrucosa) as identified from cDNA cloning experiments. Biochimica et Biophysica Acta, 1337, 1-5. GARNIER, P., GOUDEY-PERRIERE, F., BRETON, P., DEWULF, C., PETEK, F. & PERRIERE, C. 1995. Enzymatic properties of the stonefish (Synanceia verrucosa Bloch and Schneider, 1801) venom and purification of a lethal, hypotensive and cytolytic factor. Toxicon, 33, 143-55. GARNIER, P., SAUVIAT, M. P., GOUDEY-PERRIERE, F. & PERRIERE, C. 1997b. Cardiotoxicity of verrucotoxin, a protein isolated from the venom of Synanceia verrucosa. Toxicon, 35, 47-55. GEOFFROY, C. & ALOUF, J. E. 1983. Selective Purification by Thiol-Disulfide Interchange Chromatography of Alveolysin, a sulfhydryl-activated toxin of Bacillus Alvei - Toxin Properties and Interaction with Cholesterol and Liposomes. Journal of Biological Chemistry, 258, 9968-9972. GHADESSY, F. J., CHEN, D. S., KINI, R. M., CHUNG, M. C. M., JEYASEELAN, K., KHOO, H. E. & YUEN, R. 1996. Stonustoxin is a novel lethal factor from stonefish (Synanceja horrida) venom. cDNA cloning and characterization. Journal of Biological Chemistry, 271, 25575-25581. GOMES, H. L., ANDRICH, F., FORTES-DIAS, C. L., PERALES, J., TEIXEIRA-FERREIRA, A., VASSALLO, D. V., CRUZ, J. S. & FIGUEIREDO, S. G. 2013. Molecular and biochemical characterization of a cytolysin from the Scorpaena plumieri (scorpionfish) venom: Evidence of pore formation on erythrocyte cell membrane. Toxicon, 74, 92-100. GOMES, H. L., MENEZES, T. N., CARNIELLI, J. B. T., ANDRICH, F., EVANGELISTA, K. S., CHAVEZ-OLORTEGUI, C., VASSALLO, D. V. & FIGUEIREDO, S. G. 2011. Stonefish antivenom neutralises the inflammatory and cardiovascular effects induced by scorpionfish Scorpaena plumieri venom. Toxicon, 57, 992-999. GROBECKER, D. 1983. The ‘lie-in-wait’ feeding mode of a cryptic teleost, Synanceia verrucosa. In: NOAKES, D. G., LINDQUIST, D., HELFMAN, G. & WARD, J. (eds.) Predators and prey in fishes. Springer Netherlands. HAHN, S. T. & O'CONNOR, J. M. 2000. An investigation of the biological activity of bullrout (Notesthes robusta) venom. Toxicon, 38, 79-89. HALSTEAD, B. W. 1988. Poisonous and venomous marine animals of the world, Princeton, N.J., Darwin Press. HALSTEAD, B. W., CHITWOOD, M. J. & MODGLIN, F. R. 1955. The anatomy of the venom apparatus of the zebrafish, Pterois volitans (linnaeus). Anatomical Record, 122, 317-33. HALSTEAD, B. W., CHITWOOD, M. J. & MODGLIN, F. R. 1956. Stonefish Stings, and the Venom Apparatus of Synanceja horrida (Linnaeus). Transactions of the American Microscopical Society, 75, 381-397. HASHEMZADEH, M., FURUKAWA, M., GOLDSBERRY, S. & MOVAHED, M. R. 2008. Chemical structures and mode of action of intravenous glycoprotein IIb/IIIa receptor blockers: A review. Experimental & Clinical Cardiology, 13, 192-7. HERMAN, L. M. F., DEBLOCK, J. H. G. E. & MOERMANS, R. J. B. 1995. Direct-Detection of Listeria-Monocytogenes in 25 Milliliters of Raw-Milk by a 2-Step Pcr with Nested Primers. Applied and Environmental Microbiology, 61, 817-819. HOOGLAND, R., MORRIS, D. & TINBERGEN, N. 1956. The spines of sticklebacks (Gasterosteus and Pygosteus) as means of defence against predators (Perca and Esox). Behaviour, 205-236. HOPKINS, B. J., HODGSON, W. C. & SUTHERLAND, S. K. 1997. An in vitro pharmacological examination of venom from the soldierfish Gymnapistes marmoratus. Toxicon, 35, 1101-1111. HUMMON, A. B., LIM, S. R., DIFILIPPANTONIO, M. J. & RIED, T. 2007. Isolation and solubilization of proteins after TRIzol extraction of RNA and DNA from patient material following prolonged storage. Biotechniques, 42, 467-+. JHA, R. & ZI-RONG, X. 2004. Biomedical Compounds from Marine organisms. Marine Drugs, 2, 123-146. JONES, D. T., TAYLOR, W. R. & THORNTON, J. M. 1992. The rapid generation of mutation data matrices from protein sequences. Computer applications in the biosciences: CABIOS, 8, 275-282. KEHOE, M. A., MILLER, L., WALKER, J. A. & BOULNOIS, G. J. 1987. Nucleotide Sequence of the Streptolysin O (SLO) Gene: Structural Homologies between SLO and Other Membrane-Damaging, Thiol-Activated Toxins. Infection and Immunity, 55, 3228-3232. KHOO, H. E. 2002. Bioactive proteins from stonefish venom. Clinical and Experimental Pharmacology and Physiology, 29, 802-6. KHOO, H. E., CHEN, D. S. & YUEN, R. 1998. Role of free thiol groups in the biological activities of stonustoxin, a lethal factor from stonefish (Synanceja horrida) venom. Toxicon, 36, 469-476. KINI, R. M. & EVANS, H. J. 1989a. A Common Cytolytic Region in Myotoxins, Hemolysins, Cardiotoxins and Antibacterial Peptides. International Journal of Peptide and Protein Research, 34, 277-286. KINI, R. M. & EVANS, H. J. 1989b. Role of cationic residues in cytolytic activity: modification of lysine residues in the cardiotoxin from Naja nigricollis venom and correlation between cytolytic and antiplatelet activity. Biochemistry, 28, 9209-15. KIRIAKE, A. & SHIOMI, K. 2011. Some properties and cDNA cloning of proteinaceous toxins from two species of lionfish (Pterois antennata and Pterois volitans). Toxicon, 58, 494-501. KIRIAKE, A., SUZUKI, Y., NAGASHIMA, Y. & SHIOMI, K. 2013. Proteinaceous toxins from three species of scorpaeniform fish (lionfish Pterois lunulata, devil stinger Inimicus japonicus and waspfish Hypodytes rubripinnis): close similarity in properties and primary structures to stonefish toxins. Toxicon, 70, 184-93. KISHINO, H., MIYATA, T. & HASEGAWA, M. 1990. Maximum likelihood inference of protein phylogeny and the origin of chloroplasts. Journal of molecular evolution, 31, 151-160. KREGER, A. S. 1991. Detection of a cytolytic toxin in the venom of the stonefish (Synanceia trachynis). Toxicon, 29, 733-43. LEE, J. Y. L., TEOH, L. C. & LEO, S. P. M. 2004. Stonefish envenomations of the hand - A local marine hazard: A series of 8 cases and review of the literature. Annals Academy of Medicine Singapore, 33, 515-520. LIEW, H. C., KHOO, H. E., MOORE, P. K., BHATIA, M., LU, J. & MOOCHHALA, S. M. 2007. Synergism between hydrogen sulfide (H2S) and nitric oxide (NO) in vasorelaxation induced by stonustoxin (SNTX), a lethal and hypotensive protein factor isolated from stonefish Synanceja horrida venom. Life Sciences, 80, 1664-1668. LOW, K. S., GWEE, M. C., YUEN, R., GOPALAKRISHNAKONE, P. & KHOO, H. E. 1993. Stonustoxin: a highly potent endothelium-dependent vasorelaxant in the rat. Toxicon, 31, 1471-8. LU, A., YANG, L., XU, S. & WANG, C. 2014. Various Conotoxin Diversifications Revealed by a Venomic Study of Conus flavidus. Molecular & Cellular Proteomics, 13, 105-118. MALOY, W. L. & KARI, U. P. 1995. Structure-activity studies on magainins and other host defense peptides. Biopolymers, 37, 105-22. MCGIVERN, J. G. 2007. Ziconotide: a review of its pharmacology and use in the treatment of pain. Neuropsychiatric Disease and Treatment, 3, 69-85. MENEZ, A. 1998. Functional architectures of animal toxins: a clue to drug design? Toxicon, 36, 1557-72. MIYA, M., TAKESHIMA, H., ENDO, H., ISHIGURO, N. B., INOUE, J. G., MUKAI, T., SATOH, T. P., YAMAGUCHI, M., KAWAGUCHI, A., MABUCHI, K., SHIRAI, S. M. & NISHIDA, M. 2003. Major patterns of higher teleostean phylogenies: a new perspective based on 100 complete mitochondrial DNA sequences. Molecular Phylogenetics and Evolution, 26, 121-138. MOUNT, S. M. 1982. A catalogue of splice junction sequences. Nucleic Acids Research, 10, 459-72. MUSE, S. V. & GAUT, B. S. 1994. A Likelihood Approach for Comparing Synonymous and Nonsynonymous Nucleotide Substitution Rates, with Application to the Chloroplast Genome. Molecular Biology and Evolution, 11, 715-724. NAGASAKA, K., NAKAGAWA, H., SATOH, F., HOSOTANI, T., YOKOIGAWA, K., SAKAI, H., SAKURABA, H., OHSHIMA, T., SHINOHARA, M. & OHURA, K. 2009. A novel cytotoxic protein, Karatoxin, from the dorsal spines of the redfin velvetfish, Hypodytes rubripinnis. Toxin Reviews, 28, 260-265. NAKABO, T. 2000. Nihon-san gyorui kensaku : zenshu no dotei = Fishes of Japan with pictorial keys to the species, Tokyo, Tokai Daigaku Shuppankai. NEWMAN, D. J. & CRAGG, G. M. 2012. Natural Products As Sources of New Drugs over the 30 Years from 1981 to 2010. Journal of Natural Products, 75, 311-335. PARKES, D. G., MACE, K. F. & TRAUTMANN, M. E. 2013. Discovery and development of exenatide: the first antidiabetic agent to leverage the multiple benefits of the incretin hormone, GLP-1. Expert Opinion on Drug Discovery, 8, 219-44. POH, C. H., YUEN, R., KHOO, H. E., CHUNG, M., GWEE, M. & GOPALAKRISHNAKONE, P. 1991. Purification and Partial Characterization of Stonustoxin (Lethal Factor) from Synanceja Horrida Venom. Comparative Biochemistry and Physiology B-Biochemistry & Molecular Biology, 99, 793-798. REGAN, C. T. 1913. I.—The osteology and classification of the Teleostean fishes of the Order Scleroparei. The Annals and Magazine of Natural History, 11, 169-184. REIMCHEN, T. E. 1994. Predators and morphological evolution in threespine stickleback. The evolutionary biology of the threespine stickleback, 240-276. SAITOU, N. & NEI, M. 1987. The neighbor-joining method: a new method for reconstructing phylogenetic trees. Molecular Biology and Evolution, 4, 406-425. SAUNDERS, P. R. & T K S, L. 1961. Purification and properties of the lethal fraction of the venom of the stonefish Synanceja horrida (Linnaeus). Biochimica et Biophysica Acta, 52, 527-532. SAUNDERS, P. R. & TAYLOR, P. B. 1959. Venom of the lionfish Pterois volitans. American Journal of Physiology, 197, 437-40. SCARBOROUGH, R. M. 1999. Development of eptifibatide. American Heart Journal, 138, 1093-104. SCHAEFFER JR, R. C., CARLSON, R. W. & RUSSELL, F. E. 1971. Some chemical properties of the venom of the scorpionfish Scorpaena guttata. Toxicon, 9, 69-78. SCHULZ, M. H., ZERBINO, D. R., VINGRON, M. & BIRNEY, E. 2012. Oases: robust de novo RNA-seq assembly across the dynamic range of expression levels. Bioinformatics, 28, 1086-1092. SHAI, Y., HADARI, Y. R. & FINKELS, A. 1991. pH-dependent pore formation properties of pardaxin analogues. Journal of Biological Chemistry, 266, 22346-54. SHIOMI, K., HOSAKA, M., FUJITA, S., YAMANAKA, H. & KIKUCHI, T. 1989. Venoms from six species of marine fish: lethal and hemolytic activities and their neutralization by commercial stonefish antivenom. Marine Biology, 103, 285-289. SMITH, C. G. & VANE, J. R. 2003. The discovery of captopril. FASEB Journal, 17, 788-9. SMITH, W. L. & WHEELER, W. C. 2004. Polyphyly of the mail-cheeked fishes (Teleostei : Scorpaeniformes): evidence from mitochondrial and nuclear sequence data. Molecular Phylogenetics and Evolution, 32, 627-646. SMITH, W. L. & WHEELER, W. C. 2006. Venom evolution widespread in fishes: a phylogenetic road map for the bioprospecting of piscine venoms. Journal of Heredity, 97, 206-17. SWOFFORD, D. L. & DOCUMENTATION, B. 1989. Phylogenetic analysis using parsimony. Illinois Natural History Survey, Champaign. TAMURA, K., PETERSON, D., PETERSON, N., STECHER, G., NEI, M. & KUMAR, S. 2011. MEGA5: molecular evolutionary genetics analysis using maximum likelihood, evolutionary distance, and maximum parsimony methods. Molecular Biology and Evolution, 28, 2731-9. TAN, P. T., KHAN, A. M. & BRUSIC, V. 2003. Bioinformatics for venom and toxin sciences. Briefings in Bioinformatics, 4, 53-62. TERLAU, H. & OLIVERA, B. M. 2004. Conus venoms: A rich source of novel ion channel-targeted peptides. Physiological Reviews, 84, 41-68. UEDA, A., SUZUKI, M., HONMA, T., NAGAI, H., NAGASHIMA, Y. & SHIOMI, K. 2006. Purification, properties and cDNA cloning of neoverrucotoxin (neoVTX), a hemolytic lethal factor from the stonefish Synanceia verrucosa venom. Biochimica et Biophysica Acta, 1760, 1713-22. VETRANO, S. J., LEBOWITZ, J. B. & MARCUS, S. 2002. Lionfish envenomation. Journal of Emergency Medicine, 23, 379-382. WALKER, J. A., ALLEN, R. L., FALMAGNE, P., JOHNSON, M. K. & BOULNOIS, G. J. 1987. Molecular Cloning, Characterization, and Complete Nucleotide Sequence of the Gene for Pneumolysin, the Sulfhydryl-Activated Toxin of Streptococcus Pneumoniae. Infection and Immunity, 55, 1184-1189. WANG, L., SI, Y. Q., DEDOW, L. K., SHAO, Y., LIU, P. & BRUTNELL, T. P. 2011. A Low-Cost Library Construction Protocol and Data Analysis Pipeline for Illumina-Based Strand-Specific Multiplex RNA-Seq. PLoS One, 6. WARD, R. D., ZEMLAK, T. S., INNES, B. H., LAST, P. R. & HEBERT, P. D. 2005. DNA barcoding Australia's fish species. Philosophical Transactions of the Royal Society B: Biological Sciences, 360, 1847-57. YEW, W. S. & KHOO, H. E. 2000. The role of tryptophan residues in the hemolytic activity of stonustoxin, a lethal factor from stonefish (Synanceja horrida) venom. Biochimie, 82, 251-257. ZERBINO, D. R. & BIRNEY, E. 2008. Velvet: Algorithms for de novo short read assembly using de Bruijn graphs. Genome Research, 18, 821-829. | |
dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/58580 | - |
dc.description.abstract | 棘鰭魚首目中有超過1200種的魚類被認定為具有毒性。其中鮋亞目(Scorpaenoidei)為這些有毒物種的主要類群之一,因此也被認定具有高度潛力可從中發現具有生物活性之分子。在本研究中,我們解序鮋亞目中三個物種的毒素基因,並且參考資料庫已有之序列,重建出此毒素基因可能的演化過程。根據cDNA推導出來之蛋白質序列,眉鬚鱗頭鮋(Sebastapistes strongia)、尖頭擬鮋(Scorpaenopsis oxycephala)及石狗公(Sebastiscus marmoratus)毒素的兩個次單元(subunit)長度約為700個胺基酸(698~703),與過去在毒鮋屬(Synanceja)及簑鮋屬(Pterois)所發現之毒素相似。演化親緣關係的分析顯示,此毒素次單元的複製與分化應早於鮋亞目之分化,且此毒素基因可能普遍存在於整個鮋形目(Scorpaeniformes)的種類。除此之外,在簑鮋屬物種分化之前,可能發生了另外一次基因複製事件¬而產生了簑鮋屬的兩種毒素次單元。利用斑馬短鰭簑鮋(Dendrochirus zebra)的基因體,我們發現了三種不同的毒素基因序列,此結果暗示了在簑鮋亞科中假基因(pseudogene)存在的可能。此外,反轉錄定量聚合酶連鎖反應(RT-qPCR)的結果顯示,石狗公(Se. marmoratus)的背鰭硬棘具有相對最高的毒素基因表現,其他鰭的毒素表現則相對較少,而其表現量在各個不同組織的差異對應了其防禦性的功能─越容易受到攻擊的區域具有越高的毒素表現量。 | zh_TW |
dc.description.abstract | More than 1200 fish species among acanthomorpha might be venomous. Scorpaenoidei, a diverse suborder of ray-finned fish, is a hotspot of venomous fish and possesses a large pool of potential bioactive molecules. In the present study we determined the venom genes from three scorpaenoid fishes and reconstructed their evolutionary relationship. The deduced amino acid sequences of two venom subunits in Sebastapistes strongia, Scorpaenopsis oxycephala, and Sebastiscus marmoratus are about 700 a.a. (698~703 a.a.), corresponding to the size of stonefish and lionfish venoms. Phylogenetic analysis shows that the two gene subunits were duplicated prior to the speciation of Scorpaenoidei and may be common to the whole Scorpaeniform family. In addition, another gene duplication event occurred before the speciation of lionfish (Pteroinae), thus generating the two venom subunits in the genus Pterois. The venom genes determined from the genomic DNA of Dendrochirus zebra show three types of venom genes in the genome, implying that a pseudogene may present in Pteroinae. The expression pattern of the venom genes in different tissues of Se. marmoratus were evaluated by RT-qPCR, respectively. The dorsal spines were found to be the most venomous part of the fish, while lesser amounts of the venom genes were detected in other fins. The venom expression pattern seems to reflect the defensive purpose of the venom, with the most easily attacked part expressing the highest venom levels. | en |
dc.description.provenance | Made available in DSpace on 2021-06-16T08:20:46Z (GMT). No. of bitstreams: 1 ntu-103-R00241201-1.pdf: 7971981 bytes, checksum: c66002982d8728cde691b3d8b519c8e0 (MD5) Previous issue date: 2014 | en |
dc.description.tableofcontents | 中文摘要…………………………………………………………………………………I
Abstract.…………………………………………………………………………………II Table of contents………………………………………………………………………. III Table list………………………………………………………………………………..VI Figure list……………………………………………………………………………...VII Appendix list………………………………………………………………………….VIII Introduction……………………………………………………………………………...1 1.1 Applications of venom study……………………………………………….…..1 1.2 Venom studies on scorpaenoid fishes…………………………………………..2 1.3 The distribution of venom……………………………..……………………….5 1.4 Aims of this study………………………..……………………………………..6 Materials and Methods…………………………………………………………………..7 2.1 Sample collection and species identification………………...…………………7 2.2 Venom extraction and western blotting………………………………………...7 2.3 RNA and DNA extraction………………………………………………...…….9 2.3.1 RNA extraction………………………………………………………….9 2.3.2 DNA extraction………………..……………………………………….10 2.4 Reverse transcription-polymerase chain reaction (RT-PCR) and 3’ and 5’ Rapid Amplification of cDNA ends (3’ RACE and 5’ RACE)……………………….….11 2.4.1 RT-PCR……………………………………….………………………..12 2.4.2 5’ RACE-PCR……….…………………………………………………13 2.4.3 3’ RACE-PCR………………….………………………………………14 2.5 Polymerase chain reaction (PCR) and cloning……………………..…………14 2.5.1 PCR for cox1 gene……………………………………………………..14 2.5.2 PCR for the venom gene……………………………………………….15 2.5.3 PCR product cloning………………………….……………………….17 2.6 Sequence analysis………………………..……………………………………20 2.7 Phylogenic analysis…………...………………………………………………21 2.8 Reverse transcription-quantitative polymerase chain reaction (RT-qPCR)…...21 2.9. Whole genome sequencing (WGS)…………………………………………..23 Results………………………………………………………………………………….24 1. Western blotting……..……………………………………………………….24 2. Determination and differential gene expression of putative venom genes..…24 2.1 Venom genes determination from cDNA templates….………………….24 2.2 Venom genes determination from genomic DNA templates………...…..26 2.3 Open reading frames and deduced amino acid sequences analysis...……29 3. Phylogenetic analysis……………….……………………………………….30 3.1 Outgroup searching...…………………………………………………….30 3.2 Phylogenetic trees construction………………………………………….31 4. Venom gene expression pattern in Se. marmoratus………………...………..32 5. Whole genome sequencing in Se. strongia…………………………………..33 Discussion……………………………………………………………………………....36 1. High similarity among the venoms in Scorpaenoidei………………………..36 2. Conservative nature of intron positions……………..……………………….39 3. Precedence of venom gene duplication to scorpaenoid speciation….……….40 4. Defending weapons reinforcement by selection pressure…………………...44 5. Functional unit of the venom protein………………………………………..46 6. Whole genome sequencing………………….……………………………….47 Conclusion……………………………………………………………………………...48 References……………………………………………………………………………...50 Table ………………………………………………………...…………………………57 Figure…………………………………………………………………………………...65 Appendix……………………………………………………………………………….78 | |
dc.language.iso | en | |
dc.title | 鮋亞目魚類毒素基因解序及演化分析 | zh_TW |
dc.title | Venom gene determination and evolution in Scorpaenoidei | en |
dc.type | Thesis | |
dc.date.schoolyear | 102-1 | |
dc.description.degree | 碩士 | |
dc.contributor.oralexamcommittee | 黃登福(Deng-Fwu Hwang),湯森林(Sen-Lin Tang),廖德裕(Te-Yu Liao) | |
dc.subject.keyword | ?形目,?亞目,石狗公,毒素基因, | zh_TW |
dc.subject.keyword | Scorpaeniformes,Scorpaenoidei,scorpionfish,rockfish,venom gene, | en |
dc.relation.page | 93 | |
dc.rights.note | 有償授權 | |
dc.date.accepted | 2014-01-29 | |
dc.contributor.author-college | 理學院 | zh_TW |
dc.contributor.author-dept | 海洋研究所 | zh_TW |
顯示於系所單位: | 海洋研究所 |
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