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Microbial starch-binding domains (SBD) and granule-bound starch synthase I (GBSSI) are proteins which are accumulated in potato starch granules. The efficiency of SBD and GBSSI for targeting active luciferase reporter proteins to granules during starch biosynthesis was compared. GBSSI or SBD sequences were fused to the N- or C-terminus of the luciferase (LUC) gene, via an artificial Pro-Thr encoding linker sequence. The genes were introduced into an amylose-free (amf) potato mutant. It appeared that SBD was superior to GBSSI as a targeting sequence, mainly because the luciferase retained higher activity in the SBD-containing fusion proteins than in the GBSSI-containing ones. 相似文献
104.
Mutations in genes encoding melanosomal proteins cause pigmentary glaucoma in DBA/2J mice. 总被引:11,自引:0,他引:11
Michael G Anderson Richard S Smith Norman L Hawes Adriana Zabaleta Bo Chang Janey L Wiggs Simon W M John 《Nature genetics》2002,30(1):81-85
Pigmentary glaucoma is a significant cause of human blindness. Abnormally liberated iris pigment and cell debris enter the ocular drainage structures, leading to increased intraocular pressure (IOP) and glaucoma. DBA/2J (D2) mice develop a form of pigmentary glaucoma involving iris pigment dispersion (IPD) and iris stromal atrophy (ISA). Using high-resolution mapping techniques, sequencing and functional genetic tests, we show that IPD and ISA result from mutations in related genes encoding melanosomal proteins. IPD is caused by a premature stop codon mutation in the Gpnmb (GpnmbR150X) gene, as proved by the occurrence of IPD only in D2 mice that are homozygous with respect to GpnmbR150X; otherwise, similar D2 mice that are not homozygous for GpnmbR150X do not develop IPD. ISA is caused by the recessive Tyrp1b mutant allele and rescued by the transgenic introduction of wildtype Tyrp1. We hypothesize that IPD and ISA alter melanosomes, allowing toxic intermediates of pigment production to leak from melanosomes, causing iris disease and subsequent pigmentary glaucoma. This is supported by the rescue of IPD and ISA in D2 eyes with substantially decreased pigment production. These data indicate that pigment production and mutant melanosomal protein genes may contribute to human pigmentary glaucoma. The fact that hypopigmentation profoundly alleviates the D2 disease indicates that therapeutic strategies designed to decrease pigment production may be beneficial in human pigmentary glaucoma. 相似文献
105.
We have developed technologies that simplify genomic library construction and screening, substantially reducing both the time and the cost associated with traditional library screening methods and facilitating the generation of gene-targeting constructs. By taking advantage of homologous recombination in Escherichia coli, we were able to use as little as 80 bp of total sequence homology to screen for a specific gene from a genomic library in plasmid or phage form. This method, called recombination cloning (REC), takes only a few days instead of the several weeks required for traditional plaque-lift methods. In addition, because every clone in the mouse genomic library we have constructed has a negative selection marker adjacent to the genomic insert, REC screening can generate gene-targeting vectors in one step, from library screening to finished construct. Conditional targeting constructs can be generated easily with minimal additional manipulation. 相似文献
106.
Superoxide activates mitochondrial uncoupling proteins. 总被引:64,自引:0,他引:64
Karim S Echtay Damien Roussel Julie St-Pierre Mika B Jekabsons Susana Cadenas Jeff A Stuart James A Harper Stephen J Roebuck Alastair Morrison Susan Pickering John C Clapham Martin D Brand 《Nature》2002,415(6867):96-99
Uncoupling protein 1 (UCP1) diverts energy from ATP synthesis to thermogenesis in the mitochondria of brown adipose tissue by catalysing a regulated leak of protons across the inner membrane. The functions of its homologues, UCP2 and UCP3, in other tissues are debated. UCP2 and UCP3 are present at much lower abundance than UCP1, and the uncoupling with which they are associated is not significantly thermogenic. Mild uncoupling would, however, decrease the mitochondrial production of reactive oxygen species, which are important mediators of oxidative damage. Here we show that superoxide increases mitochondrial proton conductance through effects on UCP1, UCP2 and UCP3. Superoxide-induced uncoupling requires fatty acids and is inhibited by purine nucleotides. It correlates with the tissue expression of UCPs, appears in mitochondria from yeast expressing UCP1, and is absent in skeletal muscle mitochondria from UCP3 knockout mice. Our findings indicate that the interaction of superoxide with UCPs may be a mechanism for decreasing the concentrations of reactive oxygen species inside mitochondria. 相似文献
107.
胃收缩运动在消化过程中起着重要作用.传统的测量胃运动的方法是侵 袭性的.本文提出一种运用人工神经网络由体表胃电图(electrogastrogram) 无损识别胃收缩运动的方法.以5个受试者的胃电图作为训练集,另5个受试 者的胃电图作为测试集.以同时检测的与每段胃电图对应的胃腔内压力记录 作为评价标准,运用经过优化的具有单个隐层的反向传播神经网络,以分段 后的每段胃电图的时-频表征作为网络的输入,实验结果表明:识别胃运动静 止期的准确度达90呢,识别收缩运动期的准确度达94%. 相似文献
108.
Stephen K. McNees 《Journal of forecasting》1982,1(1):37-48
This article stresses how little is known about the quality, particularly the relative quality, of macroeconometric models. Most economists make a strict distinction between the quality of a model per se and the accuracy of solutions based on that model. While this distinction is valid, it leaves unanswered how to compare the‘validity’of conditional models. The standard test, the accuracy of ex post simulations, is not definitive when models with differing degrees of exogeneity are compared. In addition, it is extremely difficult to estimate the relative quantitative importance of conceptual problems of models, such as parameter instability across‘policy regimes’ In light of the difficulty in comparisons of conditional macroeconometric models, many model-builders and users assume that the best models are those that have been used to make the most accurate forecasts are those made with the best models. Forecasting experience indicates that forecasters using macroeconometric models have produced more accurate macroeconomic forecasts than either naive or sophisticated unconditional statistical models. It also suggests that judgementally adjusted forecasts have been more accurate than model-based forecasts generated mechanically. The influence of econometrically-based forecasts is now so pervasive that it is difficult to find examples of‘purely judgemental’forecasts. 相似文献
109.
Archaeopteryx, the earliest known flying bird (avialan) from the Late Jurassic period, exhibits many shared primitive characters with more basal coelurosaurian dinosaurs (the clade including all theropods more bird-like than Allosaurus), such as teeth, a long bony tail and pinnate feathers. However, Archaeopteryx possessed asymmetrical flight feathers on its wings and tail, together with a wing feather arrangement shared with modern birds. This suggests some degree of powered flight capability but, until now, little was understood about the extent to which its brain and special senses were adapted for flight. We investigated this problem by computed tomography scanning and three-dimensional reconstruction of the braincase of the London specimen of Archaeopteryx. Here we show the reconstruction of the braincase from which we derived endocasts of the brain and inner ear. These suggest that Archaeopteryx closely resembled modern birds in the dominance of the sense of vision and in the possession of expanded auditory and spatial sensory perception in the ear. We conclude that Archaeopteryx had acquired the derived neurological and structural adaptations necessary for flight. An enlarged forebrain suggests that it had also developed enhanced somatosensory integration with these special senses demanded by a lifestyle involving flying ability. 相似文献
110.