草蝦熱休克同源蛋白70之研究—基因選殖、定性及免疫功能分析

Wan-Yu Lo; 羅婉瑜

請用此 Handle URI 來引用此文件： http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/39199

完整後設資料紀錄

DC 欄位	值	語言
dc.contributor.advisor	宋延齡(Yen-Ling Song)
dc.contributor.author	Wan-Yu Lo	en
dc.contributor.author	羅婉瑜	zh_TW
dc.date.accessioned	2021-06-13T17:24:15Z	-
dc.date.available	2010-02-02
dc.date.copyright	2005-02-02
dc.date.issued	2005
dc.date.submitted	2005-01-27
dc.identifier.citation	Ali A., Salter-Cid L., Flajnik M. F. and Heikkila J. J. (1996) Isolation and characterization of a cDNA encoding a Xenopus 70kDa heat shock cognate protein, Hsc70. I. Comp. Biochem. Physiol. B Biochem. Mol. Biol. 113: 681-687. Ali K. S., Dorgai L., Abraham M. and Hermesz E. (2003) Tissue-and stressor-specific differential expression of two Hsc70 genes in carp. Biochem. Biophys. Res. Commun. 307: 503-509. Anderson J. R. (2001) The mechanisms of direct, virus-induced destruction of neurons. Curr. Top. Microbiol. Immunol. 253: 15-33. Anggraeni M. S. and Owens L. (2000) The haemocytic origin of lymphoid organ spheroid cells in the penaeid prawn Penaeus monodon. Dis. Aquat. Organ. 40(2):85-92. Arsene F., Tomoyasu T. and Bukau B. (2000) The heat shock response of Escherichia coli. Int. J. Food Microbiol. 55(1-3):3-9. Aspan A., Huang T. S., Cerenius L. and S
dc.identifier.uri	http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/39199	-
dc.description.abstract	本論文進行一系列針對草蝦熱休克同源蛋白70 (Hsc70) 的研究。此基因由草蝦血球的cDNA選殖而來，其基因全長為2207 bp，包含1959 bp之譯碼區(coding region) 及5’、3’端分別為40及208 bp的外緣區(flanking regions)，推譯此蛋白質含652個胺基酸，分子量估計約為71,481Da，等電點值約為5.2。根據演化分析，此基因應屬於本質型熱休克同源蛋白70基因 (constitutive form hsc70 )。由Native- PAGE分析發現藉由大腸桿菌表現之重組Hsc70蛋白可結合S-carboxymethyl α-lactalbumin (CMLA)，顯示此蛋白具有監護子(chaperone)的功能。在體外(in vitro) 進行ATPase分析，發現重組Hsc70蛋白可水解ATP成為ADP，且藉由結合未完整折疊胜	zh_TW
dc.description.abstract	We cloned the cDNA of the heat shock cognate 70 (hsc70) gene of tiger shrimp (Penaeus monodon). It was 2207 bp long and included a 1959 bp coding region, a 40 bp flanking region at the 5’ end and a 208 bp flanking region at the 3’end. The deduced, 652 amino acid sequence had a molecular mass of 71,481 Da and an estimated isoelectric point (pI) of 5.2. Based on phylogenetic analysis, the gene is clustered with the Hsc70 proteins of invertebrates and vertebrates. In native gel electrophoresis, recombinant P. monodon Hsc70 expressed in an E. coli system is tightly associated with S- carboxymethylated	en
dc.description.provenance	Made available in DSpace on 2021-06-13T17:24:15Z (GMT). No. of bitstreams: 1 ntu-94-D88225005-1.pdf: 4044883 bytes, checksum: bd4b11c3d3c05c605490aff36628536d (MD5) Previous issue date: 2005	en
dc.description.tableofcontents	CONTENTS 中文摘要....................................................................1 ABSTRACT....................................................................3 LITERATURE REVIEW..................................................5 Heat shock protein ........................................................5 Heat shock protein 70 family..........................................6 Mechanism of HSP70 family..........................................7 HSP70 family in viral infection.......................................8 Apoptosis in viral infection ..........................................10 HSP70 family in apoptosis ..........................................11 Characterization of white spot syndrome virus............13 Chapter I Cloning and Molecular Characterization of Heat Shock Cognate 70 (hsc70)from tiger shrimp (Penaeus monodon).........................................15 INTRODUCTION..........................................................16 MATERIALS AND METHODS......................................19 RESULTS.................................................................... 28 DISCUSSION...............................................................32 Chapter II Reduction in Hsc70 Correlates with Apoptosis in White Spot Syndrome Virus (WSSV) Infected Shrimp (Penaeus monodon)........................................36 INTRODUCTION..........................................................36 MATERIALS AND METHODS......................................41 RESULTS.....................................................................47 DISCUSSION...............................................................51 CONCLUSION.............................................................57 REFERENCES............................................................58 FIGURES AND TABLES...............................................iii APPENDIX....................................................................iv FIGURES AND TABLES Fig.1 Nucleotide and deduced amino acid sequence of P. monodon hsc70 genecDNA.................................................................................................74 Fig. 2 Phylogenetic tree showing the relationship of P. monodon hsc70 amino acid sequence to other HSP70 family members from a variety of species........................75 Fig.3 Purification of recombinant Hsc70....................................................76 Fig.4 Identification the Hsc70 / CMLA complex..........................................77 Fig.5 The effect of Hsc70 and CMLA on ATP hydrolysis............................78 Fig.6 In situ hybridization of shrimp hsc70 with antisese riboprobe...........79 Table1 Abundance of Hsc70 mRNA in P. monodon tissues as determined by in situ nucleic acid hybridization.......................................................................................80 Fig.7A Northern blot analysis of RNA extracted from P. monodon hemocytes...................................................................................................81 Fig.7B Northern blot analysis of RNA extracted from P. monodon hemocytes...................................................................................................81 Fig.8 Immuno-gold TEM of P. monodon hemocytes...................................82 Fig.9 Coimmunoprecipitation of Hsc70 and the WSSV envelop protein from hemocytes of P. monodon......................................................................................................83 Fig.10A Expression of Hsc70 in shrimp hemocytes during WSSV infection........................................................................................................84 Fig.10A Expression of Hsc70 in shrimp hemocytes during WSSV infection........................................................................................................85 Fig.10B Expression of Hsc70 in shrimp hemocytes during WSSV infection........................................................................................................86 Fig.11 Caspase-3-like protease activity in hemocytes drawn from the non-infected and WSSV-infected shrimp...................................................................................87 Fig.12 Western blot analysis of Hsc70 expression in subcuticular epithelium and gill derived from WSSV-infected shrimp...........................................................................88
dc.language.iso	en
dc.subject	監護子	zh_TW
dc.subject	細胞凋亡	zh_TW
dc.subject	草蝦	zh_TW
dc.subject	熱休克同源蛋白70	zh_TW
dc.subject	P. monodon	en
dc.subject	Apoptosis	en
dc.subject	Chaperone	en
dc.subject	Hsc70	en
dc.title	草蝦熱休克同源蛋白70之研究—基因選殖、定性及免疫功能分析	zh_TW
dc.title	Study of Heat Shock Cognate 70 in Penaeus monodon- Gene Cloning, Molecular Characterization and Immune Function Assay	en
dc.type	Thesis
dc.date.schoolyear	93-1
dc.description.degree	博士
dc.contributor.oralexamcommittee	施秀惠(Hsiu-Hui Shih),張雯(Wen Chang),陳小梨(Show-Li Chen),徐亞莉(Ya-Li Hsu),胡清華(Chin-Hwa Hu)
dc.subject.keyword	細胞凋亡,草蝦,熱休克同源蛋白70,監護子,	zh_TW
dc.subject.keyword	P. monodon,Hsc70,Chaperone,Apoptosis,	en
dc.relation.page	88
dc.rights.note	有償授權
dc.date.accepted	2005-01-27
dc.contributor.author-college	生命科學院	zh_TW
dc.contributor.author-dept	動物學研究研究所	zh_TW
顯示於系所單位：	動物學研究所

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