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研究生:傅孝瑜
研究生(外文):Tshiau-yu Fu
論文名稱:陰道鞭毛蟲紅素氧還蛋白表達研究
論文名稱(外文):Rubrerythrin, a non-haem binding protein, expresses in Trichomonas vaginalis in vitro.
指導教授:辛致煒辛致煒引用關係
指導教授(外文):Jyh-wei Shin
學位類別:碩士
校院名稱:國立成功大學
系所名稱:微生物及免疫學研究所
學門:生命科學學門
學類:微生物學類
論文種類:學術論文
論文出版年:2008
畢業學年度:96
語文別:英文
論文頁數:74
中文關鍵詞:紅素氧還蛋白陰道鞭毛蟲
外文關鍵詞:RubrerythrinTrichomonas vaginalis
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陰道鞭毛蟲 (T. vaginalis) 是一種厭氧性的單細胞原生寄生蟲,主要寄生於人類的泌尿生殖道,引起陰道鞭毛蟲症 (trichomoniasis),是最常見的非病毒的性接觸傳染疾病。流行病學研究指出感染陰道鞭毛蟲會使病人受到人類後天免疫缺乏病毒(HIV-1)感染的機會增加1.5倍。鐵藉由影響陰道鞭毛蟲的生理特性,例如:黏附因子合成、表面抗原的表達以及許多過氧化氫體(hydrogenosome)中的酵素活性,對陰道鞭毛蟲的生長與存活來說,是不可缺少的微量元素。我們利用蛋白質體學研究方法,比較在補充有12 �嵱及70 �n�嵱檸檬酸鐵(ammonium iron citrate)、12 �嵱血紅素(haemoglobin)或12 �嵱高鐵血紅素(hemin)等不同來源鐵的無鐵YI-S培養液(yeast-extract iron serum medium)中,陰道鞭毛蟲(T. vaginalis strain 30236 (JH31A, USA))的基因表達差異,並將有明顯差異的蛋白質利用胜肽質量指紋譜鑑定(peptide mass fingerprint)方法鍵定出其身份。其中,紅素氧還蛋白(rubrerythrin, TVAG_064490)是一種非血晶質(haem)鍵結的蛋白質,是一種過氧化氫酶,在檸檬酸鐵環境下有高量表現,且表現量隨著鐵濃度降低而降低,在血紅素環境下幾乎不表現,而在高鐵血紅素環境下有高量表現。RT-PCR結果顯示在mRNA表現上也有相同的情形。而在Tritrichomonas foetus和Trichomonas vaginalis strain 30001 (C-1: NIH),紅素氧還蛋白對鐵的反應也相同。與此同時,同樣具有去除氧化壓力能力的超氧岐化酶(superoxide dismutase),硫氧還蛋白過氧化酶(thioredoxin peroxidase),硫氧還蛋白(thioredoxin)及硫氧還蛋白還原酶(thioredoxin reductase)在以檸檬酸鐵或血紅素為鐵來源的培養情況下,分別有不同的基因亞型(isoform)被表現。
Trichomonas vaginalis is an anaerobic parasitic protozoan, which causes trichomoniasis, a sexually transmitted disease. T. vaginalis may play a critical and under-recognized role in amplifying HIV transmission. Epidemiological study indicates that infection with T. vaginalis increases a 1.5-fold risk of HIV-1 acquisition. Iron is an essential element for the growth and survival of T. vaginalis by regulating a variety of trichomonal properties such as adhesin synthesis, gene expression of surface immunogens and several hydrogenosomal enzyme activities of the T. vaginalis. Subproteomics analysis, we compared the gene expression of T. vaginalis strain ATCC 30236 (JH31A, USA) cultivated in the iron free yeast-extract iron serum (YIS) medium supplied with 12 �嵱 (low) or 70�n�嵱 (high) ammonium iron citrate, haemoglobin and hemin which was used for mimicking the habitant environment of T. vaginalis. A 21kDa, pI 8.16 peptide spot could be recognized 15.4-fold expression under high iron supplement than haemoglobin supplement. It was identified with peptide mass fingerprint method as rubrerythrin (RBR, TVAG_064490), a non-haem iron binding protein which was recently identified in T. vaginalis. The RT-PCR results showed RBR TVAG_064490 mRNA was regulated by iron. Not only in different strains of T. vaginalis but also in Tritrichomonas foetus and Trichomonas vaginalis strain 30001 (C-1: NIH), RBR was expressed and induced under iron supplement. At the same time, specific expression pattern of components of oxygen scavenge system such as superoxide dismutase, thioredoxin peroxidase, thioredoxin and thioredoxin reductase isoforms were induced under specific iron source condition.
Index 1
List of tables 2
List of figures 3
Abstract 5
Introduction 7
The aim of the study 13
Materials and methods 14
Results 22
Discussion 30
References 33
Tables 39
Figures 47
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