中华传染病杂志
中華傳染病雜誌
중화전염병잡지
CHINESE JOURNAL OF INFECTIOUS DISEASES
2011年
3期
134-138
,共5页
王丽丽%龚凤云%叶露%宋建新
王麗麗%龔鳳雲%葉露%宋建新
왕려려%공봉운%협로%송건신
pvdQ基因%假单胞菌,铜绿%突变%基因表达%质粒
pvdQ基因%假單胞菌,銅綠%突變%基因錶達%質粒
pvdQ기인%가단포균,동록%돌변%기인표체%질립
pvdQ gene%Pseudomonas aeruginosa%Mutation%Gene expression%Plasmids
目的 探讨铜绿假单胞菌pvdQ(PA2385)基因对群集运动的作用.方法 构建pvdQ基因表达质粒pME6032-pvdQ并鉴定,采用电穿孔法将带有pvdQ基因的质粒转入铜绿假单胞菌野生株PAO1中,构建pvdQ-高表达株.同时将空质粒pME6032采用电穿孔法转入PAO1中,构建pME6032空质粒株.选择含有反向筛选基因sacB的pEX18Gm质粒作为自杀载体,构建缺失pvdQ基因的铜绿假单胞菌无痕缺失突变株,溶菌肉汤(LB)液过夜培养,观察菌株的直径变化.统计学处理采用单因素方差分析.结果 经鉴定,成功构建pvdQ-高表达株和pvdQ-突变株,比较4株菌的群集运动直径变化:PAO1为(20.52±1.80)mm,pME6032空质粒株为(19.39±2.10)mm,pvdQ-高表达株为(51.20±2.16)mm,pvdQ-突变株为(3.30±0.55)mm.pME6032空质粒株与野生株PAO1相比,直径无显著变化(t=-0.1493,P>0.05),pvdQ-突变株与野生株PAO1相比,直径减小(t=2.8525,P<0.05),pvdQ-高表达株与野生株PAO1相比,直径增大(t=1.4230,P<0.05).结论 pvdQ基因参与调控铜绿假单胞菌群集运动,且能促进铜绿假单胞菌群集运动.
目的 探討銅綠假單胞菌pvdQ(PA2385)基因對群集運動的作用.方法 構建pvdQ基因錶達質粒pME6032-pvdQ併鑒定,採用電穿孔法將帶有pvdQ基因的質粒轉入銅綠假單胞菌野生株PAO1中,構建pvdQ-高錶達株.同時將空質粒pME6032採用電穿孔法轉入PAO1中,構建pME6032空質粒株.選擇含有反嚮篩選基因sacB的pEX18Gm質粒作為自殺載體,構建缺失pvdQ基因的銅綠假單胞菌無痕缺失突變株,溶菌肉湯(LB)液過夜培養,觀察菌株的直徑變化.統計學處理採用單因素方差分析.結果 經鑒定,成功構建pvdQ-高錶達株和pvdQ-突變株,比較4株菌的群集運動直徑變化:PAO1為(20.52±1.80)mm,pME6032空質粒株為(19.39±2.10)mm,pvdQ-高錶達株為(51.20±2.16)mm,pvdQ-突變株為(3.30±0.55)mm.pME6032空質粒株與野生株PAO1相比,直徑無顯著變化(t=-0.1493,P>0.05),pvdQ-突變株與野生株PAO1相比,直徑減小(t=2.8525,P<0.05),pvdQ-高錶達株與野生株PAO1相比,直徑增大(t=1.4230,P<0.05).結論 pvdQ基因參與調控銅綠假單胞菌群集運動,且能促進銅綠假單胞菌群集運動.
목적 탐토동록가단포균pvdQ(PA2385)기인대군집운동적작용.방법 구건pvdQ기인표체질립pME6032-pvdQ병감정,채용전천공법장대유pvdQ기인적질립전입동록가단포균야생주PAO1중,구건pvdQ-고표체주.동시장공질립pME6032채용전천공법전입PAO1중,구건pME6032공질립주.선택함유반향사선기인sacB적pEX18Gm질립작위자살재체,구건결실pvdQ기인적동록가단포균무흔결실돌변주,용균육탕(LB)액과야배양,관찰균주적직경변화.통계학처리채용단인소방차분석.결과 경감정,성공구건pvdQ-고표체주화pvdQ-돌변주,비교4주균적군집운동직경변화:PAO1위(20.52±1.80)mm,pME6032공질립주위(19.39±2.10)mm,pvdQ-고표체주위(51.20±2.16)mm,pvdQ-돌변주위(3.30±0.55)mm.pME6032공질립주여야생주PAO1상비,직경무현저변화(t=-0.1493,P>0.05),pvdQ-돌변주여야생주PAO1상비,직경감소(t=2.8525,P<0.05),pvdQ-고표체주여야생주PAO1상비,직경증대(t=1.4230,P<0.05).결론 pvdQ기인삼여조공동록가단포균군집운동,차능촉진동록가단포균군집운동.
Objective To investigate the effect of pvdQ(PA2385)gene on Pseudomonas aeruginosa swarming motility.Methods The plasmid pME6032 with pvdQ gene was constructed and identified,then transformed into Pseudomonas aeruginosa PAO1 using the eleetroporation to build pvdQ-overexpression strain.The pME6032-PAO1 strain was constructed with the same method.The cloning plasmid pEX18Gm containing sacB was successfully used to construct unmarked deletion mutant of pvdQ gene and pvdQ mutant strain. Bacteria were inoculated in LB and were cultured overnight.The clones were measured for the diameter of the swarming zone.The statistical analysis was done using one-factor ANOVA.Results Strains of pvdQ-overexpression and pvdQ-mutant were successfully constructed and confirmed by polymerase chain reaction(PCR).The four strains were compared for the swarming motility by changes in diameter:PAO1(20.52±1.80)mm,pME6032-PAO1 strain(19.39±2.10)mm,pvdQ-overexpression strain(51.20±2.16)mm,pvdQ-mutant strain(3.30±0.55)mm.The diameter of pME6032-PAO1 strain was not significantly different from that of wild strain PAO1(t=-0.1493,P>0.05).However,the diameter of pvdQ(Q-mutant strain was significantly shorter than that of wild strain PAO1(t=2.8525,P<0.05).while the diameter of pvdQ-overexpression strain was longer than that of the wild strain PAO1(t=1.4230,P<0.05).Conclusions pvdQ gene may be involved in regulating the swarming motility of Pseudomonas aeruginosa,which can promote Pseudomonas aeruginosa swarming motility.
