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1
題名:Expression and Characterization of Rice Manganese Superoxide Dismutase in Escherichia coli    
水稻超氧歧化酶在大腸桿菌中的表現及生化性質研究
著者:Yin-Chen Tzeng(曾尹貞) ;Shu-Mei Pan(潘素美)
出版地區:台灣
出版城市:台北市
學科:生命科學
刊名:Taiwania
卷期:44卷4期(1999.12)
頁碼:479-490
語言:英語
摘要: 中文摘要PDF ; 英文摘要PDF

本文報告水稻超氧歧化嗨(MnSOD) 在大腸桿菌中的表現及生化性質的分析。利 用PCR 將對應到水稻MnSOD 成熟蛋白質之正確cDNA 序列接入pGEX-4T-l 表現載 體中,將建構好的質體再轉型至大腸桿菌BL2l 品系中。融合蛋白質GST-MnSOD 可 被IPTG 誘導表現,而經由glutathione 親和膠體層析可純化均質之GST-MnSOD 融合 蛋白質。 重組之MnSOD (rMnSOD) 及GST-MnSOD 均對KCN 和H202 不敏戚,此即係有 生物中潭、態MnSOD 的一特性。兩者的單體分子量分別為23 kDa 和50 kDa '而在潭、 態上均以二元體的形式存在。rMnSOD 的等電點為4.64 ' GST-MnSOD 的等電點則介 於pH 4.74-4.975 之間。rMnSOD 及GST-MnSOD 兩蛋白質處理60 0C ' 20 分鐘後, SOD 活性下降至30% ;兩者在鹼性環境中均較穩定。
The corrected cDNA coding for rice mature MnSOD protein was made by PCR and inserted into pGEX-4T.1 expression vector. The recombinant DNA was transforrned to E. coli BL21 and expression of GST -MnSOD fusion protein was induced by addition ofIPTG to bacterial cultures. The homogeneous GST-MnSOD was purified by the one-step GST-glutathione affinity system. Both purified GST-MnSOD and recombinant MnSOD (rMnSOD) remained MnSOD activity, which showed an insensitivity towards KCN and HP2' The molecular size ofmonomer ofGST-MnSOD or rMnSOD was 50 kDa and 23 kDa respectively, and the functional forrn of both was dimer. The isoelectric point ofrMnSOD was 4.64, but that ofGST-MnSOD was in the range ofpH 4.74 - 4.97. The SOD activities ofGST-MnSOD and rMnSOD declined to 30 % after incubation at 60 Oc for 20 minutes; but they were more stable in an alkaline pH environment.


    

本卷期目次
Taiwania 44卷4期 (1999.12)
Eastern Asia as a Living Museum for Archaic Angiosperms and Other Seed Plants/ Robert F. Thorne
Eocene Angiospermous Palynomorphs of Taiwan/ Cheng-Long Shaw
Expression and Characterization of Rice Manganese Superoxide Dismutase in Escherichia coli/ Yin-Chen TzengShu-Mei Pan
Variation in Heat Shock Protein Synthesis and Genomic Diversity in the Natural Population of Aspergillus fumigatus in Northern Taiwan/ Kuei-Yu ChenZuei-Ching Chen
Notes on Puccinia Species on Cyperaceae in Taiwan/ Huang-Chi KuoZuei-Ching Chen
Two New Naturalized Grasses in Taiwan/ Chang-Sheng KuohGow-Ing LiaoMing-Yih Chen
Subcellular Localization of Calcium in the Crystal-Forming Sclereids of Nymphaea tetragona Georgi/ Ling-Long Kuo-HuangShiang-Jiuun CHEN
Sonerila Roxb. (Melastomataceae), a N ew Generic Record for the Flora of Taiwan/ Tsung-Hsin HsiehKuoh-Cheng Yang
 
   
 
   

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