重庆医科大学学报
重慶醫科大學學報
중경의과대학학보
UNIVERSITATIS SCIENTIAE MEDICINAE CHONGQING
2009年
11期
1462-1465
,共4页
袁泰先%朱兴华%吴凯峰%尹楠林%胥文春%王虹%尹一兵%张雪梅
袁泰先%硃興華%吳凱峰%尹楠林%胥文春%王虹%尹一兵%張雪梅
원태선%주흥화%오개봉%윤남림%서문춘%왕홍%윤일병%장설매
肺炎链球菌%转化%操纵子comCDE%双向凝胶电泳
肺炎鏈毬菌%轉化%操縱子comCDE%雙嚮凝膠電泳
폐염련구균%전화%조종자comCDE%쌍향응효전영
Streptococcus pneumoniae%Transformation%Operon comCDE%Two-dimensional electrophoresis
目的:探讨操纵子comCDE对调控肺炎链球菌转化的分子机制.方法:将D39野生菌株与comE基因缺陷菌株(ΔcomE)分别用感受态刺激因子(Competence-stimulating peptide,CSP)诱导转化,记录不同时间点的转化率,并采用FQ-PCR技术检测其comE的mRNA表达水平,并通过双向电泳及图像分析比较CSP诱导后不同时间点的蛋白质表达谱差异及野生菌株与ΔcomE菌株的蛋白质表达谱差异.结果:D39菌株在CSP诱导后不同时间点都能发生转化,其中0 min时转化率最高,其comE的mRNA表达量在CSP诱导10 rain后表达显著增加,20 rain时迅速回落(P＜0.05). ΔcomE菌株不能被CSP诱导发生转化.蛋白质组学研究显示,ΔcomE菌株有6个蛋白较D39菌株的表达量显著增加;CSP诱导10min时有6个蛋白表达显著增加,而在20 min时又迅速回落;二者有2个蛋白相同;有1个蛋白在加入CSP后表达显著降低.结论:基因comE的存在是CSP诱导转化必需的,CSP诱导转化可能有非comE依赖途径,而comE可能有除转化以外调控功能,且不依赖于CSP.
目的:探討操縱子comCDE對調控肺炎鏈毬菌轉化的分子機製.方法:將D39野生菌株與comE基因缺陷菌株(ΔcomE)分彆用感受態刺激因子(Competence-stimulating peptide,CSP)誘導轉化,記錄不同時間點的轉化率,併採用FQ-PCR技術檢測其comE的mRNA錶達水平,併通過雙嚮電泳及圖像分析比較CSP誘導後不同時間點的蛋白質錶達譜差異及野生菌株與ΔcomE菌株的蛋白質錶達譜差異.結果:D39菌株在CSP誘導後不同時間點都能髮生轉化,其中0 min時轉化率最高,其comE的mRNA錶達量在CSP誘導10 rain後錶達顯著增加,20 rain時迅速迴落(P＜0.05). ΔcomE菌株不能被CSP誘導髮生轉化.蛋白質組學研究顯示,ΔcomE菌株有6箇蛋白較D39菌株的錶達量顯著增加;CSP誘導10min時有6箇蛋白錶達顯著增加,而在20 min時又迅速迴落;二者有2箇蛋白相同;有1箇蛋白在加入CSP後錶達顯著降低.結論:基因comE的存在是CSP誘導轉化必需的,CSP誘導轉化可能有非comE依賴途徑,而comE可能有除轉化以外調控功能,且不依賴于CSP.
목적:탐토조종자comCDE대조공폐염련구균전화적분자궤제.방법:장D39야생균주여comE기인결함균주(ΔcomE)분별용감수태자격인자(Competence-stimulating peptide,CSP)유도전화,기록불동시간점적전화솔,병채용FQ-PCR기술검측기comE적mRNA표체수평,병통과쌍향전영급도상분석비교CSP유도후불동시간점적단백질표체보차이급야생균주여ΔcomE균주적단백질표체보차이.결과:D39균주재CSP유도후불동시간점도능발생전화,기중0 min시전화솔최고,기comE적mRNA표체량재CSP유도10 rain후표체현저증가,20 rain시신속회락(P＜0.05). ΔcomE균주불능피CSP유도발생전화.단백질조학연구현시,ΔcomE균주유6개단백교D39균주적표체량현저증가;CSP유도10min시유6개단백표체현저증가,이재20 min시우신속회락;이자유2개단백상동;유1개단백재가입CSP후표체현저강저.결론:기인comE적존재시CSP유도전화필수적,CSP유도전화가능유비comE의뢰도경,이comE가능유제전화이외조공공능,차불의뢰우CSP.
Objective: To investigate the molecular mechanism of operon comCDE regulating S.pn transformation.Methods:The wild S.pn strains and their comE-defective derivations(ΔcomE) were subjected to CSP-induced transformation.Transformation rates were calculated at different time points,and the mRNA expression levels of come were detected by FQ PCR.The proteins of different expression among different strains were detected by two-dimensional electrophoresis and image analysis.Results:D39 could undergo transformation at 0、10min and 20min after CSP was added,and the transformation rate at 0min was the highest.After 10 min utesinduction with CSP the mRNA level of comE was significantly higher than that of basal level(P＜0.05),and at 20minutes the mRNA level of comE was significantly decreased.For comE-defective strain,the transformation rate was 0.proteomics analysis displayed that six proteins were expressed significantly higher in ΔcomE strains than that in D39 wild stains.The expression of six proteins increased significantly at 10 minutes after CSP was added.but reduted quicklv at 20 minutes after CSP was added.The expression of two proteins were changedin both cases.The expression of another one protein was significantly decreased after CSP was added.Conclusion:ComE is an essential gene for CSP-induced transformation,CSP probably induces some transformation related genes by comE-independent pathway.And comE probably has other fuctions than regulated-transformation,and it might be CSP-independent.
