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91.
Joaquin Royo Isabel Diaz Pablo Rodriquez-Palenzuela Pilar Carbonero 《Plant molecular biology》1996,31(5):1051-1059
The geneItr1, encoding trypsin inhibitor BTI-CMe, has been obtained from a genomic library ofHordeum vulgare L. The gene has no introns and presents in its 5-upstream region 605 bp that are homologous to the long terminal repeats (LTR) of the copia-like retro-transposon Bare-1. Functional analysis of theItr1 promoter by transient expression in protoplasts derived from different barley tissues, has shown that in this system theItr1 promoter retains its endosperm specifity and thetrans-regulation mediated by theLys3a gene. The proximal promoter extending 343 bp upstream of the translation initiation ATG codon is sufficient to confer fullGUS expression and for endosperm specifity. In protoplasts derived from thelys3a mutant, Risø 1508,GUS activity was less than 5% of that obtained with the same constructs in the protoplasts of wild-type Bomi from which it derives. Gel retardation experiments, after incubation with proteins obtained from both types of endosperm nuclei, also show differential patterns. Possible reasons for these differences are discussed.Equal authours 相似文献
92.
Short-term aluminium uptake by tobacco cells: Growth dependence and evidence for internalization in a discrete peripheral region 总被引:1,自引:0,他引:1
Short-term uptake and initial localization of aluminium (Al) were investigated in cultured cells of Nicotiana tabacum L. cv. BY-2. Graphite furnace atomic absorption spectrometry and an in vivo Al-sensitive fluorometric assay, employing morin, yielded similar results in all experiments. Aluminium uptake was critically dependent on cell growth. As opposed to negligible uptake in stationary-phase cells, Al uptake (20 μ M AlCl3 , pH 4.5, 23°C) by actively growing cells was detectable within 5 min, with an initial rate of 16 nmol Al (106 cells)−1 h−1 . Increased CaCl2 levels (up to 20 m M ), low temperature (4°C), and pre-chelation of Al to citrate greatly reduced Al uptake (by 75–90%). A pH-associated permeabilization of cells at pH 4.5, as monitored by trypan blue, was observed in some growing cells. Although permeability to trypan blue was not a requirement for Al uptake, enhanced membrane permeability at pH 4.5, relative to pH 5.6, may contribute to Al uptake. Aluminium was observed to localize mainly in a pronounced and discrete fluorescent zone at the cell periphery (2–30 μm wide), presumably in the cortical cytosol and/or the adjoining plasma membrane section, although the possibility cannot be excluded that some Al resided in the cell wall apposing this discrete region. However, as judged by the Al-morin assay, there were no detectable Al levels in the remaining, larger portion of the cell wall. The potential of the Al-morin method in Al toxicity studies is illustrated. 相似文献
93.
Plasma membranes from the green alga Chlamydomonas reinhardtii were purified by differential centrifugation and two-phase partitioning in an aqueous polymer system. The isolated plasma membranes were virtually free from contaminating chloroplasts, mitochondria, endoplasmic reticulum and Golgi membranes as shown by marker enzyme and pigment analysis. The isolated plasma membranes exhibited vanadate sensitive ATPase activity, indicating the presence of a P-type ATPase. This was verified by using antibodies against P-type ATPase from Arabidopsis , which crossreacted with a protein of 109 kDa. The ATPase activity was inhibited to more than 90% by vanadate (Ki = 0.9 μ M ) but not affected by inhibitors specific for F- or V-type ATPases. demonstrating the purity of the plasma membranes. Mg-ATP was the substrate, and the rate of ATP-hydrolysis followed simple Michaelis-Menten kinetics giving a Km = 0.46 m M . Free Mg2+ stimulated the activity, K1/2 = 0.68 m M . Maximal activity was obtained at pH 8. The ATPase activity was latent but stimulated 10 to 20-fold in the presence of detergents. This indicates that the isolated plasma membrane vesicles were tightly sealed and mostly right-side-out, making the ATPase inaccessible to the hydrophilic substrate ATP. In the presence of the Brij 58, the isolated plasma membranes performed ATP dependent H+ -pumping as shown by the optical pH probe acridine orange. H+ -pumping was dependent on the presence of valinomycin and K+ ions and completely abolished by vanadate. Addition of Brij 58 has been shown to produce 100% sealed inside-out vesicles of plant plasma membranes (Johansson et al. 1995, Plant J. 7: 165–173) and this was also the case for plasma membranes from the green alga Chlamydomonas reinhardtii. 相似文献
94.
Kenichiro Nakashima Naotaka Kuroda Shinki Kawaguchi Mitsuhiro Wada Shuzo Akiyama 《Luminescence》1995,10(3):185-191
A sensitive peroxyoxalate chemiluminescent (PO-CL) assay for activities of oxidases (uricase, choline oxidase, cholesterol oxidase and xanthine oxidase) which catalyse a formation of hydrogen peroxide was developed using 4,4′-oxalyl-bis[(trifluoromethylsulphonyl)imino]trimethylene-bis(4-methylmorpholinium)trifluoromethanesulphonate as a chemiluminogenic reagent and 2,4,6,8-tetramorpholinopyrimido[5,4-d]pyrimidine as a fluorophore. The standard curve for hydrogen peroxide was linear over the range 1 × 10?7-1 × 10?4 mol/L. Relative standard deviations for oxidase assays were 5.1–12.7% (n = 10). Detection limits were 1 × 10?3 U/mL for uricase, 5 × 10?4 U/mL for choline oxidase, 5 × 10?3 U/mL for cholesterol oxidase and 5 × 10?4 U/mL xanthine oxidase (sample to blank ratio, 3). 相似文献
95.
The use of 6-(N-acetyl-L -phenylalanyl)-aminoluciferin as a novel substrate for α-chymotrypsin has been demonstrated. The kinetic parameters determined are KM = 0.38mmol/L, kcat = 6.5 s?1 and kcat/kM = 17,100 (L/mols). The test principle of the coupled assay is the release of aminoluciferin by enzymatic cleavage of 6-(N-acetyl-L -phenylalanyl)-aminoluciferin. Aminoluciferin is oxidized, with light emission, by firefly luciferase (Photinus pyralis) and can be quantified in a luminometric assay. The detection limit for chymotrypsin was found to be 0.3 ng per assay. 6-(N-acetyl-L -phenylalanyl)-aminoluciferin has been synthesized as an example for a new class of highly sensitive substrates. By modification of the peptide residue these new substrates may be suitable for ultrasensitive detection of different proteinases. 相似文献
96.
本文讨论了中国草蛉科已知属种的地理分布及其特点,指出:属的分布以东洋成分比重较大,高山高原属分化明显;种类分布呈现东洋成分由东部沿海向北挺进、古北种类通过西北地区向西南延伸的格局,丰富的高山、高原种类及狭布种类,是我国草蛉区系的一大特点。 相似文献
97.
戊肝病毒活性肽的选择,合成与应用 总被引:3,自引:0,他引:3
对戊型肝炎病毒(HEV)编码蛋白序列进行了亲水性分析及二级结构预测,选择亲水性强、具有β-转角与β-折叠的区段,采用多肽固相合成法合成了HEV基因组3个开读框架(ORF1,ORF2和ORF3)中可能的抗原表位,以免疫学方法进行鉴定并选出了分别来自HEV3个ORF的、具有重要生物活性与应用前景的3段肽(EH174、EH265、EH362)。在此基础上,进行了抗HEVELISA新型检测试剂盒的实验室研究及临床试用。结果表明,所研究的戊肝抗体检测试剂盒特异性高、临床符合性好、具有可重复性,在戊肝辅助诊断及流行病学调查中具有重要意义。 相似文献
98.
激光诱变选育AC10菜用大青豆的研究报告 总被引:2,自引:1,他引:1
采用有性杂交和激光红宝石辐照交替进行,经10多年的研究、试验、选育出一个蛋白质含量、脂肪含量特高的菜用大青豆-AC10。AC10菜用大青豆,系早熟、适应性广、抗逆性强、耐迟播、产量高、效益好的品种。全生育期110天、80-85天采摘青毛豆、单产鲜毛豆700公斤以上,老豆单产160公斤左右。据农业部谷物测试中心分析,蛋白质含量48.32%,脂肪含量21.36%,合计69.68%。查新结果表明,为国内 相似文献
99.
七星瓢虫(Coccmella septimpunctata)为了适应环境的变化,通过咽侧体产生保幼激素的活动调节其生殖作用。为了探索内外因素对咽侧体活动的影响,应用放射化学法及免疫电泳测定了食物、卵巢发育、脑神经肽、保幼激素类似物对卵黄发生期成虫保幼激素生物合成及血淋巴中卵黄原蛋白含量的影响。结果表明咽侧体活性受上述各种因子的影响。咽侧体活性与卵黄原蛋白含量及卵母细胞生长密切相关,说明有反馈作用。食物的质与量影响着咽侧体活性的变化。低剂量外源保幼激素类似物处理成虫则可促使咽侧体活性的变化。脑分泌的神经肽(allatotropin)可活化咽侧体。这些结果表明雌瓢虫保幼激素的生产主要是受起源于脑的促咽侧体信号的调节作用。 相似文献
100.
We present here microwave-based modifications of standard protein assays that dramatically reduce the time required to determine
protein concentrations. Typical protein determinations involve incubation times ranging from 15–60 min. Microwave irradiation
of specimens reduces this time requirement to 10–20 s without compromising accuracy or reliability. The remarkable speed with
which protein determinations may be carried out using microwave enhancement greatly simplifies general laboratory procedures
that depend on the estimation of protein concentrations.
An erratum to this article is available at . 相似文献