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標題: 雞傳染性鼻炎菌莢膜合成基因之特性分析
Characterization of The Capsule Biosynthesis Genes from Avibacterium paragallinarum
作者: 羅義凱
Lo, Yi-Kai
關鍵字: Avibacterium paragallinarum
出版社: 微生物暨公共衛生學研究所
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摘要: 雞傳染性鼻炎菌 (Avibacterium paragallinarum) 為雞傳染性可利查病之病原菌,又稱雞傳染性鼻炎病,會引起雞急性上呼吸道疾病,受感染的雞隻會出現發燒、流鼻水、眼窩下竇腫脹、發出囉音等症狀,也造成經濟上的損害,包括種雞與肉雞生長遲緩、蛋雞生蛋率降低等等。莢膜為細菌致病力之主要決定因子之一,目前許多研究報告皆集中雞傳染性鼻炎致病因子的探討與相關有效疫苗的研製,但是莢膜相關的報告資料甚少。故本論文在探討雞傳染性鼻炎菌的莢膜合成基因,結果發現了標準株H18莢膜合成基因中除了含有ccbEF1F2CB之基因序列外,還存在有acbEDGHCB之基因序列。序列比對發現,ccbEF1F2CB與Pasteurella multocida type D之dcbEFCB基因以及E. coli K5之kfiABCD基因相似度高,而dcbEFCB與kfiABCD等基因所表現之蛋白皆為與合成heparosan相關之酵素,故推論為H18莢膜菌株中含有heparosan。另一方面,H18之acbEDGHCB基因與Pasteurella multocida type F之fcbEDCB基因最為相似,而fcbEDCB基因所表現之蛋白為合成chondroitin相關之酵素,推論H18莢膜菌株中含有chondroitin。因此H18莢膜中可能同時存在heparosan與chondroitin。為瞭解H18中ccbEF1F2CB與acbEDGHCB等基因在細菌染色體中之所在位置,本研究利用PCR與gene-walking技術對上述基因進行分析,結果發現ccbEF1F2CB基因位於莢膜合成基因之region I與region III之間,而acbEDGHCB之上游為region III,而下游位置則仍待鑑定。本研究為首度發現Av. paragallinarum H18之莢膜中可能同時存在heparosan與chondroitin兩種成份,以及其基因體中具有兩個region III。本研究也利用RT-PCR及Western blot技術來進行H18菌株中ccbEF1F2CB與acbEDGHCB之基因表現分析。
Avibacterium paragallinarum is the causative agent of infectious coryza (IC), an acute upper respiratory tract disease of chickens. Infected chickens usually show fever, nasal mucus, eyehole edema, rale, diarrhea. Coryza causes significant economic losses in poultry industry, including retardation of growth in breeders and broilers and a drop in egg production in layers. The aim of this study was to investigate the capsule biosynthesis genes of Avibacterium paragallinarum strain H18, which is a reference strain of serovar C. The results showed that the capsule biosynthesis genes of strain H18 comprised two gene clusters, ccbEF1F2CB and acbEDGHCB. Sequence comparison showed that ccbEF1F2CB genes were similar to dcbEFCB genes from Pasteurella multocida type D and kfiABCD genes from E. coli K5. Because proteins encoded by dcbEFCB and kfiABCD genes are involved in the biosynthesis of heparosan, it is possible that the capsule of strain H18 contains heparosan. On the other hand, because the acbEDGHCB genes are similar to fcbEDCB genes from P. multocida type F, and proteins encoded by fcbEDCB genes are involved in the biosynthesis of chondroitin, it is possible that the capsule of strain H18 contains chondroitin. Therefore, both heparosan and chondroitin are present in the capsule of strain H18. To investigate chromosomal location of ccbEF1F2CB genes and acbEDGHCB genes in strain H18, we conducted PCR and gene-walking analyses of ccbEF1F2CB and acbEDGHCB. The result suggested that ccbEF1F2CB genes are located between the region I and region III of the capsule biosynthesis gene cluster. In contrast, the upstream sequences of acbEDGHCB genes are region III, whereas the downstream sequences of acbEDGHCB genes remain unknown. This is the first report showing that capsule of strain H18 contains heparosan and chondroitin and that two region III sequences are present in the genomes of Avibacterium paragallinarum strain H18.
其他識別: U0005-2007201010425900
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