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121.
This article presents for the first time a modified protocol for RNase protection analysis that allows the substitution of32P with33P without loss of the high sensitivity of this method achieved with32P. With this protocol, we were able to detect at least 1 pg of specific mRNA. In the RNase protection analysis33P labeled riboprobes are more advantageous with regard to an easier handling and better resolution. 相似文献
122.
在吲哚乙酸不同位点偶联载体蛋白对其抗体特异性的影响 总被引:2,自引:0,他引:2
分别选择吲哚乙醇分子上的C1位羧基和吲哚环上的N位作为偶联载体蛋白的位点,用混合酸酐法和甲醛搭桥法分别合成了两种免疫原IAA—CO—NH一HSA和IAA-N-BSA,并进而制得了对吲哚乙酸侧链识别能力不同的两种多克隆抗体,分别可特异识别甲酯化IAA和游离态IAA;用碳化二亚胺法和甲醛搭桥法分别合成IAA—CO—NH-BSAbIAA—N—OVA两种复合物,以之为包被物,建立了两种IAAELISA。其灵敏度分别为0.35pmol和1.80ppmol;检测范围分别为0.78~800pmol和1.95~2000pmol;批内变异系数分别为4.45%和4.79%;批间变异系数分别为1.15%和1.50%。笔者用这两种ELISA检测了兰花气生根和桑树苗样品中IAA的含量,发现两种检测结果相当一致。 相似文献
123.
Intercellular signaling molecules of the transforming growth factor- (TGF-) superfamily are required for pattern formation in many multicellular organisms. The decapentaplegic (dpp) gene of Drosophila melanogaster has several developmental roles. To improve our understanding of the evolutionary diversification of this large family we identified dpp in the grasshopper Schistocerca americana. S. americana diverged from D. melanogaster approximately 350 million years ago, utilizes a distinct developmental program, and has a 60-fold-larger genome than D. melanogaster. Our analyses indicate a single dpp locus in D. melanogaster and S. americana, suggesting that dpp copy number does not correlate with increasing genome size. Another TGF- superfamily member, the D. melanogaster gene 60A, is also present in only one copy in each species. Comparison of homologous sequences from D. melanogaster, S. americana, and H. sapiens, representing roughly 900 million years of evolutionary distance, reveals significant constraint on sequence divergence for both dpp and 60A. In the signaling portion of the dpp protein, the amino acid identity between these species exceeds 74%. Our results for the TGF- superfamily are consistent with current hypotheses describing gene duplication and diversification as a frequent response to high levels of selective pressure on individual family members. 相似文献
124.
125.
Ken Lee Akihiro Ito †Kunio Koshimura †Tetsuya Ohue †Yasutaka Takagi †Soichi Miwa 《Journal of neurochemistry》1995,64(2):874-882
Abstract: Hypoxia is known to disturb neuronal signal transmission at the synapse. Presynaptically, hypoxia is reported to suppress the release of neurotransmitters, but its postsynaptic effects, especially on the function of neurotransmitter receptors, have not yet been elucidated. To clarify the postsynaptic effects, we used cultured bovine adrenal chromaffin cells as a model of postsynaptic neurons and examined specific binding of l -[3 H]nicotine (an agonist for nicotinic acetylcholine receptors: nAChRs) and 22 Na+ flux under control and hypoxic conditions. Experiments were performed in media preequilibrated with a gas mixture of either 21% O2 /79% N2 (control) or 100% N2 (hypoxia). Scatchard analysis of the specific binding to the cells revealed that the KD under hypoxic conditions was twice as large as that under control conditions, whereas the B max was unchanged. When the specific [3 H]nicotine binding was kinetically analyzed, the association constant ( k 1 ) but not the dissociation constant ( k −1 ) was decreased to 40% of the control value by hypoxia. When the binding assay was performed using the membrane fraction, these changes were not observed. Nicotine-evoked 22 Na+ flux into the cells was suppressed by hypoxia. In contrast, specific [3 H]quinuclidinyl benzilate binding to the intact cells was unaffected by hypoxia. These results demonstrate that hypoxia specifically suppresses the function of nAChRs (and hence, neuronal signal transmission through nAChRs), primarily by acting intracellularly. 相似文献
126.
Craig J. Coates Catherine L. Turney Marianne Frommer David A. O'Brochta W. D. Warren Peter W. Atkinson 《Molecular & general genetics : MGG》1995,249(2):246-252
Plasmid-based excision assays performed in embryos of two non-drosophilid species using the mariner transposable element from Drosophila mauritiana resulted in empty excision sites identical to those observed after the excision of mariner from D. mauritiana chromosomes. In the presence of the autonomous mariner element Mos1, excision products were recovered from D. melanogaster, D. mauritiana and the blowfly Lucilia cuprina. When a hsp82 heat shock promoter-Mos1 construct was used to supply mariner transposase, excision products were also recovered from the Queensland fruitfly Bactrocera tryoni. Analysis of DNA sequences at empty excision sites led us to hypothesise that the mariner excision/repair process involves the formation of a heteroduplex at the excision breakpoint. The success of these assays suggests that they will provide a valuable tool for assessing the ability of mariner and mariner-like elements to function in non-drosophilid insects and for investigating the basic mechanisms of mariner excision and repair. 相似文献
127.
The Sea of Marmara, an intercontinental basin with shallow and narrowstraits, connects the Black and Mediterranean Seas. Data
obtained during1991–1996 have permitted the determination of the elementalcomposition of seston in the euphotic zone and the
N:P ratio of thesubhalocline waters of the Marmara Sea. Since primary production is alwayslimited to the less saline upper
layer (15–20 m), of the Marmara Sea,the subhalocline waters of Mediteranean origin are always rich in nutrients(NO3 + NO2 = 8–10 μm, PO4 = 0.8–1.2 μm) but depleted in dissolvedoxygen (30–50 μm) throughout the basin, yielding an -O_2 : N : P ratio of 178 : 9 : 1. Pollution of the surfacewaters since the 60s has modified the subhalocline nutrient chemistryslightly.
In the euphotic zone, the N : P ratio of the seston changes from5.9 to 9.5 between the less and more productive periods. Though
the biologyof the Marmara has changed significantly during the previous two decades,the close relationship observed between
the elemental composition of thesurface seston and the NO3 : PO4 ratio of thesubhalocline waters strongly suggests that during the whole year primaryproduction throughout the basin and POM
export to the lower layer remainnitrogen-limited. This suggestion needs to be confirmed by bio-assays,biological studies and
sediment trap data from the upper subhaloclinedepths. Nonetheless, the counterflows in the Marmara basin possessrelatively
low N : P ratios in both dissolved and particulate nutrients andextend as far as the adjacent seas.
This revised version was published online in July 2006 with corrections to the Cover Date. 相似文献
128.
Expression of betaine aldehyde dehydrogenase gene and salinity tolerance in rice transgenic plants 总被引:9,自引:0,他引:9
Betaine as one of osmolytes plays an important role in osmoregulation of most high plants. Betaine aldehyde dehydrogenase C BADH) is the second enzyme involved in betaine biosynthesis. The BADH gene from a halophite, Atriplex hortensis, was transformed into rice cultivars by bombarment method. Totally 192 transgenic rice plants were obtained and most of them had higher salt tolerance than controls. Among transgenic plants transplanted in the saline pool containing 0.5% NaCl in a greenhouse, 22 survived, 13 of which set seeds, and the frequency of seed setting was very low, only 10% . But the controls could not grow under the same condition. The results of BADH ac-tivity assay and Northern blot showed that the BADH gene was integrated into chromosomes of transgenic plants and expressed. 相似文献
129.
高必需氨基酸转基因马铃薯的研究 总被引:8,自引:0,他引:8
80年代以来,马铃薯遗传转化系统日趋成熟,转基因工程植株已被广泛应用于基础科学研究[1]。作为食物蛋白和能量主要来源的马铃薯,提高其蛋白质含量及质量的遗传工程研究正受到人们的普遍关注[2]。Yang等[2]将旨在改善氨基酸平衡的CAT-HEAAE(氯酶素乙酰转移酶-高含量人体必需氨基酸)融合基因导入马铃薯,获得了Southernblot、Northernblot、Westernblot的证据,但尚缺少氨基酸分析的资料。玉米醇溶蛋白(zein)[3]是一个富含甲硫氨酸的贮存蛋白,它和人工合成的HE… 相似文献
130.