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

Study of Heat Shock Cognate 70 in Penaeus monodon- Gene Cloning, Molecular Characterization and Immune Function Assay

羅婉瑜(Wan-Yu Lo)

指導教授 ： 宋延齡

國立臺灣大學/生命科學院/動物學研究研究所/博士(2005年)

https://doi.org/10.6342/NTU.2005.01691

摘要

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

關鍵字

細胞凋亡 ； 草蝦 ； 熱休克同源蛋白70 ； 監護子

並列摘要

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

並列關鍵字

P. monodon ； Hsc70 ； Chaperone ； Apoptosis

參考文獻

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