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Title: | 腸炎弧菌小型核糖核酸 RyhB 的調控 Characterization of the regulatory roles of small RNA RyhB in Vibrio parahaemolyticus |
Authors: | Hao-Yu Weng 翁浩瑜 |
Advisor: | 李佳音(Chia-Yin Lee) |
Keyword: | 小型核糖核酸,腸炎弧菌, small RNA,RyhB,vibrio parahaemolyticus, |
Publication Year : | 2014 |
Degree: | 碩士 |
Abstract: | RyhB 是一個被廣泛研究的小型核糖核酸,可與標的 mRNA 鹼基配對進行調控目標基因,進而影響細菌生理及致病力。本研究以腸炎弧菌 Vibrio parahaemolyticus no.93 作為實驗菌株,建構腸炎弧菌 ryhB 刪除突變株及補償株,探討腸炎弧菌中,鐵離子相關基因、化學趨化性相關基因、移動性相關基因以及毒性因子相關基因的表現是否亦受到 RyhB 的調控影響。本篇研究證實腸炎弧菌 V. parahamoelyticus no.93 當中之小型核糖核酸 RyhB,經由 qRT-PCR 及北方墨點法分析基因轉錄表現,結果顯示與鐵離子平衡相關的超氧歧化酶 sodB (VP2118)、ferric coprogen 外膜受體 fhuE (VPA0150),化學趨化性相關之趨化性蛋白 cheY (VP2331),移動性相關的鞭毛驅動蛋白 motA (VPA1556)、 motB (VP0690)、σ54-依賴調節因子 lafK (VPA1538)、鞭毛基因 flaD (VP0790)、極性鞭毛蛋白 flaK (VP2253) 及鞭毛蛋白 fliS (VP2254),毒性相關基因中的吸附因子 mam7 (VP1611)、T3SS1 調節子 toxS (VP0819) 及 toxR (VP0820) 皆受到 RyhB 負調控而抑制其表現,iscS (VP0596) 受到 RyhB 正調控而促進其表現,而 rpoS (VP2553) 不受 RyhB 的調控。而於補償株中觀察到 sdhC (VP0843)、vopQ (VP1680)、vopS (VP1686)、VPA0450、phospholipase C (VP1098) 五個基因未能回復其野生株的現象。推測除了 RyhB 外,尚有因子影響基因之表現。本研究證實 RyhB 在腸炎弧菌中扮演的調控角色,除了與鐵離子攝取外,亦與化學趨化性、移動性及 T3SS1 之調控相關。 RyhB, one of the well-studied sRNAs, exert their regulatory function by short base pairing with their target mRNAs. In this study, adopting Vibrio parahaemolyticus no. 93 to construct the deletion mutant to investigate iron-related genes, chemotaxis and stress factor genes, motility genes and virulence genes in Vibrio parahaemolyticus. Resulting of qRT-PCR and northern blotting indicate that the iron-related genes of superoxide dismutase sodB (VP2118), outer membrane receptor for ferric coprogen fhuE(VPA0150), chemotaxis-related gene of chemotaxis protein cheY(VP2331), motility-related genes of flagella motor protein motA(VPA1556), motB(VP0690),σ54-dependent regulator lafK (VPA1538), flagellin flaD (VP0790), polar flagellar protein flaK (VP2253) and fliS (VP2254), virulence genes of adhesion factor mam7 (VP1611), type 3 secretion system 1 (T3SS1) regulator toxS (VP0819) and toxR (VP0820) negative regulated by RyhB. iscS (VP0596) positive regulated by RyhB. rpoS (VP2553) is not regulated by RyhB. By observing complementary strain, the result indicate that sdhC (VP0843), vopQ (VP1680), vopS (VP1686),VPA0450 and phospholipase C (VP1098) can not rescue their gene expression in wild type strain, showing that besides RyhB, there is other factor which can effect gene expression. In this study, confirmed that besides of iron-related genes, chemotaxis genes, motility genes and virulence genes also can be regulted by RyhB. |
URI: | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/55827 |
Fulltext Rights: | 有償授權 |
Appears in Collections: | 農業化學系 |
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ntu-103-1.pdf Restricted Access | 2.23 MB | Adobe PDF |
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