摘要
6-磷酸果糖激酶是糖酵解途径中的关键酶,它催化糖酵解途径中第一个不可逆反应。本研究利用pK18mobsacB自杀质粒采用同源双交换的方法对野油菜黄单胞菌Xcc8004中的6-磷酸果糖激酶基因(XC_0872)进行缺失突变,获得无标记的缺失突变体DM0872。表型检测结果显示DM0872突变体不影响野油菜黄单胞菌对葡萄糖和果糖的利用,不影响胞外多糖的合成,也不影响其致病性。该结果显示糖酵解途径在野油菜黄单胞菌的地位并不重要。另外,我们利用RT-PCR方法检测了XC_0872的转录情况,结果显示XC_0872在Xcc8004中是转录的。而之前曾有报道称黄单胞菌中无法检测出6-磷酸果糖激酶活性,这表明XC_0872进行了转录后调控从而使6-磷酸果糖激酶活性受到限制。本研究为野油菜黄单胞菌中糖酵解途径的调控提供了理论依据,对揭示野油菜黄单胞菌中该途径的调控机制具有一定的意义。
6-phosphofructokinase which catalyzes the first irreversible reaction in the glycolytic pathway,is a well-known key enzyme of glycolysis.In order to identify the function of the 6-phosphofructokinase gene（XC_0872） in Xcc8004 strain,XC_0872 was deleted using the suicide plasmid pK18mobsacB,and the marker-free deletion mutant DM0872 was obtained.The mutant analysis showed that DM0872 had the same ability to aquire glucose or fructose as Xcc8004 strain and produced EPS as much as the wild-type strain.DM0872 also performed almost the same virulence level as Xcc8004.These may indicate glycolysis is not very important to Xcc.According some previous reports glycolysis was supposed to be absent in Xcc,as no 6-phosphofructokinase activity had been observed.Our RT-PCR result showed that the transcription of XC_0872 was organized in Xcc.So there must be some unknown regulations to limited the activity of 6-phosphofructokinase after transcription.This research would provide the theory basis for the regulatory of glycolysis pathway in Xcc,which may be significant to reveal the regulatory mechanism of that pathway.
出处
《基因组学与应用生物学》
CAS
CSCD
北大核心
2010年第6期1047-1054,共8页
Genomics and Applied Biology
