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201.
Characterization of the interaction of the glp repressor of Escherichia coli K-12 with single and tandem glp operator variants. 总被引:2,自引:2,他引:0
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N Zhao W Oh D Trybul K S Thrasher T J Kingsbury T J Larson 《Journal of bacteriology》1994,176(8):2393-2397
The glp operons of Escherichia coli are negatively controlled by the glp repressor. Comparison of the repressor-binding affinities for consensus and altered consensus operators in vivo showed that all base substitutions at positions 3, 4, 5, and 8 from the center of the palindromic operator caused a striking decrease in repressor binding. Substitutions at other positions had a severe to no effect on repressor binding, depending on the base substitution. The results obtained indicate that the repressor binds with highest affinity to operators with the half-site WATKYTCGWW, where W is A or T, K is G or T, and Y is C or T. Strong cooperative binding of the repressor to tandem operators was demonstrated in vivo. Cooperativity was maximal when two 20-bp operators were directly repeated or when 2 bp separated the two operators. Cooperativity decreased with the deletion of 2 bp or the addition of 4 bp between the individual operators. Cooperativity was eliminated with a 6-bp insertion between the operators. 相似文献
202.
Kelem Kassahun Pei Hu Mark P. Grillo Margaret R. Davis Lixia Jin Thomas A. Baillie 《Chemico-biological interactions》1994,90(3):253-275
The ability of 2-n-propyl-4-pentenoic acid (Δ4-VPA) and 2-n-propyl-2(E)-pentenoic acid ([E]-Δ2-VPA), two unsaturated metabolites of valproic acid (VPA), to form reactive intermediates, deplete hepatic glutathione (GSH) and cause accumulation of liver triglycerides was investigated in the rat. With the aid of ionspray liquid chromatography-tandem mass spectrometry (LC-MS/MS), three GSH adducts were detected in the bile of Δ4-VPA-treated animals and were identified as 4-hydroxy-5-glutathion-S-yl-VPA-γ-lactone, 5-glutathion-S-yl-(E)-Δ3-VPA and 3-oxo-5-glutathion-S-yl-VPA. A fourth conjugate was identified tentatively as 4-glutathion-S-yl-5-hydroxy-VPA. Quantitative analysis of the corresponding N-acetylcysteine (NAC) conjugates in urine indicated that metabolism of Δ4-VPA via the GSH-dependent pathways accounted for approximately 20% of an acute dose (100 mg kg−1 i.p.). In contrast, when rats were given an equivalent dose of (E)-Δ2-VPA, only one GSH adduct (5-glutathion-S-yl-(E)-Δ3-VPA) was detected at low concentrations in bile. In vitro experiments with rat liver mitochondria demonstrated that Δ4-VPA undergoes coenzyme A- and ATP-dependent metabolic activation in this organelle via the β-oxidation pathway to intermediates which bind covalently to proteins. When liver homogenates and hepatic mitochondria from rats injected with Δ4-VPA, (E)-Δ2-VPA or VPA were analyzed for GSH content, it was found that only Δ4-VPA depleted GSH pools significantly. Treatment of rats with Δ4-VPA and (to a lesser extent) VPA led to an accumulation of liver triglycerides, whereas (E)-Δ2-VPA had no measurable effect. It is concluded that Δ4-VPA undergoes metabolic activation by both microsomal cytochrome P-450-dependent and mitochondrial coenzyme A-dependent processes, and that the resulting electrophilic intermediates, which are trapped in part by GSH, may mediate the hepatotoxic effects of this compound. In contrast, (E)-Δ2-VPA is not transformed to any appreciable extent to reactive metabolites, which thus accounts for the apparent lack of hepatotoxicity of this positional isomer in the rat. 相似文献
203.
The entire mitochondrial genome of Schizosaccharomyces pombe ura4-294h
-was analyzed by the 2D pulsed field gel electrophoresis technique developed by Brewer and Fangman. The genome consists of multimers with an average size of 100 kb and analysis of the overlapping restriction fragments of the complete mitochondrial DNA (mtDNA) genome resulted in simply Y 2D gel patterns. Large single-stranded DNA molecules or double-stranded DNA molecules containing large or numerous single-stranded regions were found in the S. pombe mtDNA preparation. The replication of mtDNA monomers was found to occur in either direction. On the basis of these results, a replication mechanism for S. pombe mtDNA that is most consistent with a rolling circle model is suggested. 相似文献
204.
Factors affecting viability of IVF-derived bovine blastocysts after freezing and thawing were investigated. A total of 1,101 ova matured and fertilized in vitro were cultured under 2 different conditions, 1) in TCM-199 on granulosa cell monolayers at 5% CO(2) in air and 2) in synthetic oviduct fluid (SOF) medium without somatic cell support at 5% CO(2), 5% O(2), 90% N(2). All blastocysts that developed from the 2 different culture systems were individually classified into 4 grades of embryo quality and were then frozen by conventional slow freezing. Developmental rates of the IVF-derived ova to blastocysts and the survival rates of the frozen-thawed blastocysts were not different between the SOF medium (16 and 49%) and the co-culture system (13 and 61%, respectively). Survival of frozen-thawed blastocysts was affected by embryo quality in both the SOF and co-culture systems (P<0.001). Blastocysts produced in vitro were also individually classified into 3 developmental stages and were then cultured for 3 d in the co-culture system with granulosa cells after freezing and thawing. There was a difference in the survival rate of frozen-thawed embryos between blastocyst developmental stages (early vs mid, P<0.05; mid vs expanded, P<0.01; early vs expanded, P<0.001). The post-thawing survival rate of blastocysts frozen at Day 7 (62%) of culture was higher compared with that of Day 8 (45%), but there was no difference in survival rate between Day 7 and 8 of culture. The results indicate that the quality and developmental stage of blastocysts are important factors influencing their survival after freezing and thawing. 相似文献
205.
Jin Feng Kunitoshi Yamanaka Hironori Niki Teru Ogura Sota Hiraga 《Molecular & general genetics : MGG》1994,243(2):136-147
The nucleotide sequence was determined of the region upstream of the mukB gene of Escherichia coli. Two new genes were found, designated kicA and kicB (killing of cell); the gene order is kicB-kicA-mukB. Promoter activities were detected in the regions immediately upstream of kicB and kicA, but not in front of mukB. Gene disruption experiments revealed that the kicA disruptant was nonviable, but the kicB-disrupted mutant and the mutant lacking both the kicB and kicA genes were able to grow. When kicA disruptant cells bearing a temperature-sensitive replication plasmid carrying the kicA
+ gene were grown at 30° C and then transferred to 42° C, the mutant cells gradually lost colony-forming ability, even in the presence of a mukB
+ plasmid. Rates of protein synthesis, but not of RNA or DNA synthesis, fell dramatically during incubation at 42° C. These results suggested that the kicB gene encodes a killing factor and the kicA gene codes for a protein that suppresses the killing function of the kicB gene product. It was also demonstrated that KicA and KicB can function as a post-segregational killing system, when the genes are transferred from the E. coli chromosome onto a plasmid. 相似文献
206.
Summary To isolate a novel gene that contains an SH2 domain, we devised a rapid and nonradioactive cDNA library screening method using polymerase chain reaction (PCR). For PCR amplification, we designed degenerate oligonucleotide primers from the multialigned DNA sequences of SH2 domains. This method offers an inexpensive and efficient approach for the isolation of clones of interest from cDNA libraries. 相似文献
207.
IN THIS STUDY IT IS REPORTED THAT: (1) the levels of blood platelet-activating factor and serum tumour necrosis factor significantly increased after coronary ligation and reperfusion, compared with sham-ligated controls, in an anaesthetized rat model; (2) compared with vehicle controls, pretreatment with the PAF antagonist BN 50739 (10 mg/kg, i.v.) produced significant decreases in infarct size (from 29.6 +/- 4.0% to 22.4 +/- 2.1%, p < 0.05 after 3 h ligation, and from 28.5 +/- 9.5% to 10.5 +/- 4.5%, p < 0.01 after 4 h reperfusion) and the level of serum TNF (from 10.4 +/- 7.7 U/ml to 3.9 +/- 4.8 U/ml, p < 0.05); and (3) a significan positive correlation was found between the level of blood PAF or serum TNF and infarct size. The present results indicate that PAF and TNF may be important mediators involved in myocardial ischaemia and reperfusion injury, and that PAF antagonists may exert a protective effect on ischaemic or reperfused myocardium by inhibiting the interaction of PAF and TNF. 相似文献
208.
209.
三褶脉紫菀中的新二萜甙 总被引:1,自引:0,他引:1
三褶脉紫菀(AsterageratoidesTurcz.)系菊科多年生草本植物,遍布全国,是民间常用的中药,有清热解毒、祛痰镇咳的功效[1,2]。化学工作者们曾从其同属植物紫菀(AstertararicusL.f.)中分离到紫菀酮(shionone)、槲皮素(quercetin)、无羁萜(friedelin)、表无羁萜(epifriedelinel)、毛叶醇(lachnophyllol)、乙酸毛叶酯(lachnophyllolacetate)、茴香醚(anethole)以及紫菀三萜皂甙[2—4]… 相似文献
210.
向日葵幼苗环旋运动的三维轨迹 总被引:1,自引:0,他引:1
采用光学显微镜标尺和垂线原理制作的三维空间点测定仪,对向日葵(Helianthus annuusL.)幼苗的环旋运动进行了连续测量。结果表明:向日葵环旋运动的轨迹有椭圆型、摆动型和不规则型;同一植株在不同生长阶段所表现的环旋运动轨迹不一定相同,同一株龄的不同个体也不一定具有相同的运动轨迹;运动的方向有左旋和右旋,圆周运动光源可以显著地改变运动方向;从三维角度看,在整个下胚轴生长阶段,环旋运动的振幅存在一个由小变大再由大变小的变化规律 相似文献