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171.
Pollen Fertility Restoration by Nuclear Gene Fr in Cms Bean: Nuclear-Directed Alteration of a Mitochondrial Population 总被引:4,自引:0,他引:4
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Two nuclear genes, Fr and Fr2, have been identified that restore pollen fertility to cytoplasmic male sterile (CMS) common bean (Phaseolus vulgaris L.) by apparently distinct mechanisms. Whereas Fr2 appears to suppress the expression of a male sterility associated mitochondrial sequence (designated pvs), Fr restores pollen fertility by causing the elimination of this unusual mitochondrial DNA segment. To further investigate the mechanism of Fr action, Fr and Fr2 were cointroduced into the nucleus of a bean line containing the sterility inducing cytoplasm. When the effect of pvs was suppressed by Fr2, the presence of Fr no longer directed the elimination of the mitochondrial pvs sequence. This result suggests that the Fr function is dependent on proper expression of the pvs sequence. To evaluate the temporal and spatial patterns of Fr action, we undertook a polymerase chain reaction-based approach to trace the fate of the pvs sequence in different tissues of F(2) and F(3) fertile-restored plants derived from a genetic cross between a cytoplasmic male sterile line of common bean, CMS-Sprite (frfr), and fertility restorer line R351 (FrFr). We demonstrate that the Fr-directed disappearance of pvs sequence occurs during flower development. Elimination of the pvs sequence from developing megaspores results in permanent fertility restoration in the following generations. Genetic analysis demonstrated that permanent fertility restoration, that is, the complete elimination of pvs from reproductive tissues requires two doses of the Fr allele or the absence of fr in F(2) individuals. The effect of Fr was reversible until full fertility was achieved. On the basis of these results, we propose a model for the mechanism of pvs elimination by the Fr gene and discuss the dynamics of pvs-containing mitochondrial transmission in the presence of the Fr gene. 相似文献
172.
本文采用Y染色体特异的性别决定基因(Sry)作为新的细胞遗传标志,通过PCR技术来追踪观察造血干细胞的增殖与分化性能。该方法具有简便、灵敏和特异等优点。雌性受体小鼠输注雄鼠骨髓细胞和13天脾结节(CFU-S13)细胞后,Sry PCR测试受体小鼠的CFU-S结果表明,它们均为供体来源的XY细胞。用Sry PCR骨髓细胞和骨髓中脾结节生成细胞(CPU-S)的长期重建造血能力,结果表明,在存活雌性小鼠 相似文献
173.
甘蔗和烟草幼叶原生质体的核酸含量与核酸酶活性及其影响因素 总被引:1,自引:0,他引:1
与幼叶组织相比,酶法新鲜分离的甘蔗和烟草幼叶原生质体内的RNA、DNA及总核酸含量均降低。其原因可能是刚游离的原生质体内酸性和碱性RNA酶与DNA酶等活性提高所致。甘蔗叶原生质体内的核酸降低量和RNA酶与DNA酶活性的增加程度均高于烟草。随用作渗透压稳定剂的甘露醇浓度增加,甘蔗和烟草叶原生质体的RNA酶和DNA酶活性均相应提高。其中以甘蔗叶原生质体的核酸酶活性增加水平较明显。在细胞壁降解产物的作用下,除了甘蔗原生质体内的RNA酶活性略被促进外,其DNA酶和烟草叶原生质体内的核酸酶均不受影响 相似文献
174.
175.
Rotavirus RNA polymerase requires the core shell protein to synthesize the double-stranded RNA genome. 总被引:11,自引:6,他引:5
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Rotavirus cores contain the double-stranded RNA (dsRNA) genome, RNA polymerase VP1, and guanylyltransferase VP3 and are enclosed within a lattice formed by the RNA-binding protein VP2. Analysis of baculovirus-expressed core-like particles (CLPs) has shown that VP1 and VP2 assemble into the simplest core-like structures with replicase activity and that VP1, but not VP3, is essential for replicase activity. To further define the role of VP1 and VP2 in the synthesis of dsRNA from viral mRNA, recombinant baculoviruses containing gene 1 (rBVg1) and gene 2 (rBVg2) of SA11 rotavirus were generated and used to express recombinant VP1 (rVP1) and rVP2, respectively. After purification, the proteins were assayed individually and together for the ability to catalyze the synthesis of dsRNA in a cell-free replication system. The results showed that dsRNA was synthesized only in assays containing rVP1 and rVP2, thus establishing that both proteins are essential for replicase activity. Even in assays containing a primer-linked mRNA template, neither rVP1 nor rVP2 alone directed RNA synthesis. Characterization of the cis-acting replication signals in mRNA recognized by the replicase of rVP1 and rVP2 showed that they were the same as those recognized by the replicase of virion-derived cores, thus excluding a role for VP3 in recognition of the mRNA template by the replicase. Analysis of RNA-protein interactions indicated that the mRNA template binds strongly to VP2 in replicase assays but that the majority of the dsRNA product neither is packaged nor stably associates with VP2. The results of replicase assays performed with mutant VP2 containing a deletion in its RNA-binding domain suggests that the essential role for VP2 in replication is linked to the protein's ability to bind the mRNA template for minus-strand synthesis. 相似文献
176.
帕里红景天的化学成分研究 总被引:10,自引:0,他引:10
从帕里红景天根茎的石油醚和乙醇提取部分共分得14种结晶性化合物,经光谱分析和化学反应,分别鉴定为二十二醇、二十六酸、十九醇、β-谷甾醇、二十九醇、红景天甙、麦芽糖、棉皮素-8-葡萄糖甙、胡萝卜甙、酪醇、咖啡酸、没食子酸、形花内酯和新化合物帕里甙。 相似文献
177.
He Liu H. Peter Spielmann Nikolai B. Ulyanov David E. Wemmer Thomas L. James 《Journal of biomolecular NMR》1995,6(4):390-402
Summary The effect of experimental and integration errors on the calculations in interproton distances from NOE intensities is examined. It is shown that NOE intensity errors can have a large impact on the distances determined. When multiple spin (spin diffusion) effects are significant, the calculated distances are often underestimated, even when using a complete relaxation matrix analysis. In this case, the bias of distances to smaller values is due to the random errors in the NOE intensities. We show here that accurate upper and lower bounds of the distances can be obtained if the intensity errors are properly accounted for in the complete relaxation matrix calculations, specifically the MARDIGRAS algorithm. The basic MARDIGRAS algorithm has been previously described [Borgias, B.A. and James, T.L. (1990) J. Magn. Reson., 87, 475–487]. It has been shown to provide reasonably good interproton distance bounds, but experimental errors can compromise the quality of the resulting restraints, especially for weak cross peaks. In a new approach introduced here, termed RANDMARDI (random error MARDIGRAS), errors due to random noise and integration errors are mimicked by the addition of random numbers from within a specified range to each input intensity. Interproton distances are then calculated for the modified intensity set using MARDIGRAS. The distribution of distances that define the upper and lower distance bounds is obtained by using N randomly modified intensity sets. RANDMARDI has been used in the solution structure determination of the interstrand cross-link (XL) formed between 4-hydroxymethyl-4,5,8-trimethylpsoralen (HMT) and the DNA oligomer d(5-GCGTACGC-3)2 [Spielmann, H.P. et al. (1995) Biochemistry, 34, 12937–12953]. RANDMARDI generates accurate distance bounds from the experimental NOESY cross-peak intensities for the fixed (known) interproton distances in XL. This provides an independent internal check for the ability of RANDMARDI to accurately fit the experimental data. The XL structure determined using RANDMARDI-generated restrains is in good agreement with other biophysical data that indicate that there is no bend introduced into the DNA by the cross-link. In contrast, isolated spin-pair approximation calculations give distance restraints that, when applied in a restrained molecular dynamics protocol, produce a bent structure.Abbreviations NOE
nuclear Overhauser effect
- SD
standard deviation
- HMT
4-hydroxymethyl-4,5,8-trimethylpsoralen
- XL
psoralen-DNA interstrand cross-link 相似文献
178.
179.
肝细胞生长因子mRNA在成年小鼠和人胎儿不同器官中的表达 总被引:2,自引:0,他引:2
本实验从成年小鼠和胎龄4-5月的人胎儿不同器官中分离总RNA。经斑点印迹分析显示,肝细胞生长因子(HGF)mRNA在成年KM小鼠多种器官中表达,其表达水平由高到低依次为:肺、肝、肾、卵巢、睾丸、大脑和胃;在脾、心、骨髓、小肠和骨骼肌组织中以HGFmRNA。在胎龄4-5月的人胎儿中,HGFmRNA表达水平由高到低依次为:大脑、肝、腮腺、胃、小肠、肾、心和骨骼肌;肺和脾组织为阴性。由此可见,HGF在成 相似文献
180.
大鼠颈上神经节烟碱电流的整流及失敏 总被引:1,自引:0,他引:1
在培养的新生大鼠颈上神经节交感神经元标本上,用全细胞膜片钳技术研究了烟碱型乙酰胆碱受体(nAChR)通道的整流及失敏现象。烟碱激动剂引起的全细胞电流在膜电位为负值时随膜电位呈线性改变,而在膜电位达+60mV时仍不出现外向电流,表现出强烈的内向整流。nAChR通道电流存在失敏现象,即持续恒压喷射激动剂所引起的全细胞电流随时间呈指数衰减,不能保持在峰值水平,失敏随激剂浓度呈量效关系,膜电位的超极化也加 相似文献