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991.
从耐热碱性磷酸酶(TAP)200多个随机突变体的克隆库中选出耐热性明显下降的4株突变体,进行全序列及表达产物的最高耐受温度和最适反应温度测定和酶分子高级结构的模拟,分析突变位点、高级结构和耐热性表现三者的关系,探讨引起耐热性变化的机理。结构模拟显示所有突变位点都仅能引起细微的、局部的结构变化,除T320→I外都未直接触及酶的活性中心;结构上的细微改变虽然对最适反应温度影响不明显,但却使最高耐受温度降低了10℃左右;T320→I靠近酶的活性中心,尽管未能引起结构的较大变化,但却使最高耐受温度和最适反应温度同时显著降低。可见,多数点突变对高级结构的影响都不剧烈,但对耐热性尤其是最高耐受温度的影响却比较明显,一般地,在非活性区的突变通常只能引起最高耐受温度的降低,靠近活性区的突变则能同时引起最适反应温度和最高耐受温度的降低。 相似文献
992.
Joan Villanova Antonio Cano Alfonso Albacete Alfonso López Emilio Á. Cano Manuel Acosta José Manuel Pérez-Pérez 《Plant Growth Regulation》2017,81(3):511-521
Efficient propagation of uniform starting material is a critical requirement for mass production of most ornamental plants, including carnation. For some elite cultivars, the production of young plantlets is limited by poor adventitious root formation from stem cuttings. We previously characterized the molecular signature during adventitious rooting in two carnation cultivars, 2101-02 MFR and 2003 R 8, which were selected because of their contrasting rooting performance. To determine additional factors that contribute to the differences observed in adventitious rooting during the commercial scaling-up of this species, we characterized rooting performance and endogenous hormone levels in stem cuttings of these two cultivars during one production season. We found that stem cutting production declined during the harvest season in a cultivar-dependent manner. In addition, the initiation of adventitious roots in the stem cutting base depended on its endogenous auxin and cytokinin levels at harvest time, while their subsequent growth and development was mainly influenced by the physiological status of the mother plant at harvest time and of the stem cutting during the rooting process. 相似文献
993.
Ruxandra F. Sîrbulescu Iulian Ilieş Günther K. H. Zupanc 《Journal of comparative physiology. A, Neuroethology, sensory, neural, and behavioral physiology》2009,195(7):699-714
In contrast to mammals, teleost fish exhibit an enormous potential to regenerate adult spinal cord tissue after injury. However,
the mechanisms mediating this ability are largely unknown. Here, we analyzed the major processes underlying structural and
functional regeneration after amputation of the caudal portion of the spinal cord in Apteronotus leptorhynchus, a weakly electric teleost. After a transient wave of apoptotic cell death, cell proliferation started to increase 5 days
after the lesion and persisted at high levels for at least 50 days. New cells differentiated into neurons, glia, and ependymal
cells. Retrograde tract tracing revealed axonal re-growth and innervation of the regenerate. Functional regeneration was demonstrated
by recovery of the amplitude of the electric organ discharge, a behavior generated by spinal motoneurons. Computer simulations
indicated that the observed rates of apoptotic cell death and cell proliferation can adequately explain the re-growth of the
spinal cord.
Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users. 相似文献
994.
马拉硫磷是一种高效低毒的有机磷杀虫剂, 分子量大且结构特殊, 广泛用于农业害虫的防治。羧酸酯酶突变是昆虫对有机磷类杀虫剂产生代谢抗性的重要机制之一。本实验室前期已从棉蚜Aphis gossypii、 褐飞虱Nilaparvata lugens、 斜纹夜蛾Spodoptera litura、 家蚕Bombyx mori、 异色瓢虫Harmonia axyridis、 赤拟谷盗Tribolium castaneum和西方蜜蜂Apis mellifera中各克隆了一个非特异性羧酸酯酶基因, 通过体外定点突变构建了G/A151D和W271L两种突变体, 并进行了原核细胞表达和纯化。本实验在体外测定了这7种昆虫野生型和两种突变型羧酸酯酶对马拉硫磷的降解。结果显示: 棉蚜、 西方蜜蜂、 斜纹夜蛾、 赤拟谷盗的野生型羧酸酯酶能够降解马拉硫磷, 两个突变并不能提高它们的降解活性, 而家蚕、 异色瓢虫和褐飞虱的野生型羧酸酯酶不能降解马拉硫磷, G/A151D和/或W271L突变能使这些酯酶获得马拉硫磷羧酸酯酶(MCE)的活性, 有可能使这些昆虫对马拉硫磷产生抗性。不同物种的MCE活性相差较大, 斜纹夜蛾的MCE活性最高, 其kcat/Km值为1.8~1.9 L/μmol·min, 其次是赤拟谷盗, 其Kcat/Km值为0.87~0.95 L/μmol·min, 其他昆虫的MCE活性相对较低, 相差可高达10倍。 相似文献
995.
996.
昆虫嗅觉感受细胞电反应计算机数据分析 总被引:3,自引:0,他引:3
昆虫嗅觉感受细胞电反应计算机数据分析吴锋,吴才宏(北京志同电子公司北京100080)(北京大学生命科学学院北京100871)昆虫触角电位和单个嗅觉感受细胞电反应记录已成为研究昆虫性信息素及化学通讯机制的重要手段。实验中要记录和分析处理大量数据。在所记... 相似文献
997.
成体多能干细胞,如来自骨髓和脂肪组织的间充质干细胞等具有多向分化的潜能。虽然自体干细胞移植已经发展成为器官移植的有效代替疗法之一,但是由于移植位点细胞的流失和分化条件的限制等问题使得这种疗法的效率大大降低。本研究目的是将由脂肪干细胞分化而来的类肝细胞制备成具有稳定细胞性状的可移植的肝细胞片。首先在体外分离扩增脂肪干细胞,并通过控制严格地分化条件获得类肝细胞。然后将此细胞接种到聚N-异丙基丙烯酰胺(PNIPAAm)结合的细胞培养皿表面,通过调节培养温度到20oC,使细胞成片脱离培养皿形成细胞片。对细胞片进行了常规HE染色和免疫组化观察,结果显示:这类细胞片中平均含有2~3层细胞,并且保持了细胞外基质的完整。同传统的胰酶消化收集移植用细胞相比,细胞片方法极大地减少了对移植用细胞的细胞膜和细胞外基质的损伤,这将大大促进细胞片和原位组织的相互作用,增加细胞利用效率,从而有望提高治疗效果。 相似文献
998.
随着质谱技术的快速发展,蛋白质组学已成为继基因组学、转录组学之后的又一研究热点,寻找可靠的差异表达蛋白对于生物标记物的发现至关重要.因此,如何准确、灵敏地筛选出差异蛋白已成为基于质谱的定量蛋白质组学的主要研究内容之一.目前,针对该问题的研究方法众多,但这些方法策略的适用范围不尽相同.总体来说,基于质谱技术筛选差异蛋白的统计学策略可以分为3类:基于经典统计学派的策略、基于贝叶斯学派的统计检验策略和其他策略,这3类方法有各自的应用范围、特点及不足.此外,筛选过程还将产生部分假阳性结果,可以采用其他方法对差异表达蛋白的质量进行控制,以提高统计检验结果的可靠性. 相似文献
999.
Can Baysal Luisa Bortesi Changfu Zhu Gemma Farré Stefan Schillberg Paul Christou 《Molecular breeding : new strategies in plant improvement》2016,36(8):108
Genome editing with the CRISPR/Cas9 system allows mutations to be induced at any 20-bp target site in the genome preceded by the short protospacer adjacent motif (PAM) 5′-NGG-3′. The brevity and degeneracy of the PAM ensures that the motif occurs every ~10 bp in plant genomes, and all plant genes therefore contain many targetable sites. However, the CRISPR/Cas9 system tolerates up to three mismatches in the target site, so the ability to target genes in a specific manner requires the design of synthetic guide RNAs (sgRNAs) that do not bind off-target sites anywhere else in the genome. This is straightforward for single-copy genes but more challenging if a target gene has one or more paralogs because the principles that balance targeting efficiency (the frequency of on-target mutations) and accuracy (the absence of off-target mutations) are not fully understood and may be partially species-dependent. To investigate this phenomenon in rice, we targeted the rice starch branching enzyme IIb gene (OsBEIIb) with two sgRNAs designed to differ at two and six positions, respectively, from corresponding sites in the close paralog OsBEIIa. In each case, half of the mismatches were in the essential seed region immediately upstream of the PAM, where exact pairing is thought to be necessary, and the other half were in the distal part of the target. The sgRNAs also differed in predicted targeting efficiency (39 and 96 %, respectively). We found that the sgRNA with the low predicted efficiency was actually the most efficient in practice, achieving a mutation frequency of 5 % at the target site, whereas the sgRNA with the high predicted efficiency generated no mutations at the second target site. Furthermore, neither of the sgRNAs induced an off-target mutation in the OsBEIIa gene. Our data indicate that efficiency predictions should be tested empirically because they do not always reflect the experimental outcome and that a 1-bp mismatch in the seed region of a sgRNA is sufficient to avoid off-target effects even in closely related rice genes. 相似文献
1000.
Ana O. Fagundes Maira R. Aguiar Claudia S. Aguiar Giselli Scaini Monique U. Sachet Nayara M. Bernhardt Gislaine T. Rezin Samira S. Valvassori João Quevedo Emilio L. Streck 《Neurochemical research》2010,35(11):1675-1680
Methylphenidate is commonly used for the treatment of attention deficit/hyperactivity disorder. There are still few works
regarding the effects of methylphenidate on brain energy metabolism. Thus, in the present study we evaluated the effect of
chronic administration of methylphenidate on the activities of mitochondrial respiratory chain complexes I and III in the
brain of young rats. The effect of acute administration of methylphenidate on mitochondrial respiratory chain complexes I,
II, III and IV in the brain of young rats was also investigated. For acute administration, a single injection of methylphenidate
was given to rats on postnatal day 25. For chronic administration, methylphenidate injections were given starting at postnatal
day 25 once daily for 28 days. Our results showed that complexes I and III were not affected by chronic administration of
methylphenidate. Moreover, the acute administration of methylphenidate decreased complex I activity in cerebellum and prefrontal
cortex, whereas complexes II, III and IV were not altered. 相似文献