New Fluorescent Techniques Applied to σ Functions.

所有细菌都有主要的sigma因子，使RNA聚合酶能够识别其最大的启动子，而C.D.170是E的主要sigma因子。coli. The largest finding in this res\is the aggregation of D170 in res\to environment such as high physiological temperature and the end of log phase。The purified C170 C170也在类似的温度下产生聚合，同时a mutant protein with aaa 130-374存在于所有温度下的混合物和聚合物。Monomeric mutant蛋白质是活跃的，作为野生类型的C-D170-D1在纯化的重新构建的转录系统中，而oligomers在活动中占大多数的15%。Thus the domain of a130 -374 is not essential for transcManagement reaction but has essential role to prevent aggregation of Oracle D170。We have first succeeded to constructed a disruptant of rpoD （the gene of生物D170生物D1） and the mutant生物D170生物D1 and SigA， the major sigma of B。subtilis lacking aa 130-374，补充了有限条件中的破坏性，并提供aa 130-374在体内不需要最低限度的转录。通过清除Cayi D170-D1与羟基激进分子的暴露表面，Cayi D170 Cayi D1已被确认为单体和聚合形式。在聚合上保护的区域Sandwich在聚合上暴露的区域，建议超过两个相互作用的站点的存在，我们已经确定了欧米茄子单位的作用。大肠杆菌RNA聚合酶。The core enzyme prepared from omega-depleted cells binds GroEL shaperonin， and the removal of GroEL destroys the ability of core enzyme to bind D170。这证明了欧米茄的作用是成熟的核心酶，这解释了为什么欧米茄的依赖不会带来任何形式的现象。这证明了欧米茄的作用是成熟的核心酶，这解释了为什么欧米茄的依赖不会带来任何形式的现象。

All bacteria have major sigma factors, which enable RNA polymerase to recognize most of their promoters, and σィイD170ィエD1 is the major sigma factor of E. coli. The largest finding in this research is the aggregation of σィイD170ィエD1 in response to environment such as high physiological temperature and the end of log phase. The purified σィイD170ィエD1 also makes aggregates at similar temperature, while a mutant protein with deletion of aa 130-374 exists as a mixture of monomer and aggregates at all temperatures. The monomeric mutant protein is as active as the wild-type σィイD170ィエD1 in purified reconstituted transcription system, while oligomers have at most 15% of the activity. Thus the domain of aa 130-374 is not essential for transcription reaction but has essential role to prevent aggregation of σィイD170ィエD1. We have first succeeded to constructed a disruptant of rpoD (the gene of σィイD170ィエD1) and the mutant σィイD170ィエD1 and SigA, the major sigma of B. subtilis lacking aa 130-374, complement the disruption in limited conditions, proving that aa 130-374 is not required for minimal transcription in vivo.By cleaving σィイD170ィエD1 with hydroxide radical, the exposed surfaces of σィイD170ィエD1 were identified in monomeric and aggregated forms. The regions protected upon aggregation sandwich the regions exposed upon aggregation, suggesting the existence of more than two interacting sites for aggregation.We have determined the role of omega subunit of E. coli RNA polymerase. The core enzyme prepared from omega-depleted cells binds GroEL shaperonin, and the removal of GroEL destroys the ability of core enzyme to bind σィイD170ィエD1. This proves the role of omega is to maturate core enzyme, and explain why the depletion of omega does not bring any phenotype. This proves the role of omega is to maturate core enzyme, and explain why the depletion of omega does not bring any phenotype.