锌指核酸酶技术在植物基因工程中的应用

锌指核酸酶(zinc finger nucleases,ZFNs)由3到4个锌指结构(zinc fingers,ZFs)和FokⅠ核酸内切酶的剪切结构域组成。锌指核酸酶(ZFNs)通过锌指结构(ZFs)与特异核酸位点结合,再利用FokⅠ的酶切作用切割DNA,引起特异位点DNA双链断裂(double strand break,DSB)。DNA双链断裂可以通过非同源末端连接(non-homologous end joining,NHEJ) 或同源重组(homologous recombination,HR)来修复。在修复过程中实现对基因组DNA的靶向修饰。介绍了锌指核酸酶结构、人工构建途径,作用机理和试验步骤,重点综述了锌指核酸酶技术在植物基因工程的应用。     

Serum zinc levels and Alzheimer’s disease

Concentrations of zinc in postmortem serum and four brain regions were measured by flame atomic absorption spectrometry and instrumental neutron activation analysis, respectively, in nine Alzheimer’s disease (AD) and eight control subjects. A statistically significant elevation of zinc serum was observed in AD subjects (136.4±66.8 μg/dL) compared with age-matched control subjects (71.1±35.0 μg/dL). No significant differences were observed between AD and control zinc concentrations in the amygdala, hippocampus, cerebellum, and superior and middle temporal gryi.     

利用人工锌指蛋白核酸酶进行植物基因定点突变和置换

基因定点突变技术在基因组原位改变基因特定序列,避免常规转基因过程中位置效应和插入失活。定点突变生物体不含转基因或标记基因,降低风险性。高等植物基因定点突变研究初见端倪,将可能为基因原位功能研究、作物遗传改良和分子设计提供有效策略。利用锌指蛋白核酸酶（Zinc Finger Nucleases, ZFN）引入DNA定点断裂（Double-Strand Breaks, DSBs）可以高效介导基因定点突变,使得ZFN在基因定点突变中倍受关注。文章综述了植物基因定点突变的一般策略,重点介绍了锌指蛋白的结构、原理、应用,特别是ZFN介导的植物基因定点突变与置换研究进展,并对ZFN介导的植物基因定点突变与置换应用前景进行了讨论。     

宁夏贺兰山东麓荒漠草原区地衣的物种多样性研究

地衣是宁夏贺兰山东麓荒漠草原区一类重要的生物资源,地衣结皮的形成为该区域景观的稳定作出了重要的贡献。该研究于2014~2019年在贺兰山东麓荒漠草原区开展了地衣的野外调查,利用表型特征和DNA序列分析且对采集的500余份标本进行了物种的鉴定,并分析了其物种组成及地理成分。结果显示：(1)共鉴定地衣19科41属74种,其中中国新记录25种,新种2个(待发表),新记录属11个,宁夏新记录56种。(2)研究区地衣物种区系地理成分以北温带成分为主(33.78%),其他主要区系成分包括欧亚成分和世界广布成分等,具有明显的北温带成分区系特征。(3)研究区地衣以壳状和鳞状地衣为主(81.1%),生长型体现了地衣对当地气候条件的适应特性。实地调查发现宁夏贺兰山东麓荒漠草原区地衣的生存环境非常脆弱。该研究结果有助于提高对该区域地衣物种多样性的认识并促进其保护。     

新疆地衣研究现状与展望

地衣在自然界中是一种特殊的生物类群,具有独特的结构特征和较强的环境适应能力。它在环境污染评价,药物和香料开发利用方面具有较高的研究价值。目前对地衣的研究涉及地衣分类、地衣区系地理、地衣群落生态学、地衣化学、利用地衣评价环境质量以及地衣分子生物学等领域。回顾了新疆地衣研究近20年的发展概况,总结了新疆地衣研究的发展历程、特点,并展望了新疆地衣研究的方向。     

地衣标本的采集、制作与保存

详细论述了地衣标本的采集、制作和保存方法及注意事项,同时阐明了在采集标本的同时进行共生菌和光合共生物分离培养的重要性。     

The zinc/thiolate redox biochemistry of metallothionein and the control of zinc ion fluctuations in cell signaling

Free zinc ions are potent effectors of proteins. Their tightly controlled fluctuations ("zinc signals") in the picomolar range of concentrations modulate cellular signaling pathways. Sulfur (cysteine) donors generate redox-active coordination environments in proteins for the redox-inert zinc ion and make it possible for redox signals to induce zinc signals. Amplitudes of zinc signals are determined by the cellular zinc buffering capacity, which itself is redox-sensitive. In part by interfering with zinc and redox buffering, reactive species, drugs, toxins, and metal ions can elicit zinc signals that initiate physiological and pathobiochemical changes or lead to cellular injury when free zinc ions are sustained at higher concentrations. These interactions establish redox-inert zinc as an important factor in redox signaling. At the center of zinc/redox signaling are the zinc/thiolate clusters of metallothionein. They can transduce zinc and redox signals and thereby attenuate or amplify these signals.     

硫酸羟氯喹口服辅助治疗糜烂型口腔扁平苔癣的效果及对AT-Ⅲ、ZAG的影响

摘要 目的：探讨硫酸羟氯喹口服辅助治疗糜烂型口腔扁平苔癣的效果及对人抗凝血酶Ⅲ（AT-Ⅲ）、锌α2糖蛋白（ZAG）的影响。方法：选择2018年12月-2021年1月在我院接受治疗的115例糜烂型口腔扁平苔癣患者,采用随机数表法分为治疗组（n=58）和对照组（n=57）。对照组给复方倍他米松治疗,治疗组在对照组的基础上联合硫酸羟氯喹口服治疗。比较两组临床疗效、AT-Ⅲ、ZAG、白细胞介素4（IL-4）、白细胞介素17（IL-17）、干扰素γ（IFN-γ）、糜烂面积、疼痛评分水平变化情况及不良反应发生情况。结果：治疗后,两组总有效率比较差异显著（P<0.05）;治疗前,治疗组和对照组血清AT-Ⅲ、ZAG比较无显著差异;治疗后,治疗组和对照组血清AT-Ⅲ、ZAG均随着时间的推移而降低,且治疗组均低于对照组,差异显著（P<0.05）;治疗前,治疗组和对照组实验室指标水平比较无显著差异;治疗后,治疗组和对照组IL-4、IL-17均随着时间的推移而降低,且治疗组均低于对照组,IFN-γ均随着时间的推移而升高,且治疗组均高于对照组,差异显著（P<0.05）;治疗前,治疗组和对照组糜烂面积、疼痛评分比较无显著差异;治疗后,治疗组和对照组糜烂面积、疼痛评分均随着时间的推移而降低,且治疗组均低于对照组,差异显著（P<0.05）;两组不良反应总发生率为3.45%、8.77%,无显著差异（P>0.05）。结论：在糜烂型口腔扁平苔癣中应用硫酸羟氯喹口服辅助治疗疗效显著,可有效改善患者AT-Ⅲ、ZAG水平。     

植物蛋白oleosin及其在植物基因工程中的应用

Oleosin是一类特殊贮藏蛋白,主要在植物种子特异表达,覆盖于油体表面,在油体发生到分解消失过程中起着重要的生物学作用,在植物基因工程研究中有重要的应用价值。介绍oleosin的分类、分布、基本组成、结构与功能、oleosin基因及其表达,讨论oleosin在植物基因工程中的应用 。     

电子计算机在植物学中的应用

电子计算机技术的广泛应用和蓬勃发展是本世纪科学技术的卓越成就之一,它将逐渐成为各项科学技术现代化的一个重要标志。电子计算机从1946年问世以来,在近四十年内迅速经历了三代更新。目前正处在第     

Photoactivated perylenequinone toxins in fungal pathogenesis of plants

Several genera of plant pathogenic fungi produce photoactivated perylenequinone toxins involved in pathogenesis of their hosts. These toxins are photosensitizers, absorbing light energy and generating reactive oxygen species that damage the membranes of the host cells. Studies with toxin-deficient mutants and on the involvement of light in symptom development have documented the importance of these toxins in successful pathogenesis of plants. This review focuses on the well studied perylenequinone toxin, cercosporin, produced by species in the genus Cercospora. Significant progress has been made recently on the biosynthetic pathway of cercosporin, with the characterization of genes encoding a polyketide synthase and a major facilitator superfamily transporter, representing the first and last steps of the biosynthetic pathway, as well as important regulatory genes. In addition, the resistance of Cercospora fungi to cercosporin and to the singlet oxygen that it generates has led to the use of these fungi as models for understanding cellular resistance to photosensitizers and singlet oxygen. These studies have shown that resistance is complex, and have documented a role for transporters, transient reductive detoxification, and quenchers in cercosporin resistance.     

Zinc Supplementation or Regulation of its Homeostasis: Advantages and Threats

To accomplish its multifunctional biological roles, zinc requires precise homeostatic mechanisms. There are efficient mechanisms that regulate zinc absorption from the alimentary tract and its excretion by the kidney depending on the organism demands. The regulatory mechanisms of cellular zinc inflow, distribution, and zinc outflow are so efficient that symptoms of zinc deficiency are rare, and symptoms connected with its massive accumulation are even more rare. The efficiency of homeostatic mechanisms that prevent zinc deficiency or excessive zinc accumulation in the organism is genetically conditioned. It seems that an essential element of zinc homeostasis is the efficiency of zinc transmembrane exchange mechanisms. Intracellular free zinc concentration is higher than in extracellular space. Physiologically, the active outflow of zinc ions from the cell depends on the increase of its concentration in extracellular space. The ion pumps activity depends on the efficiency by which the cell manages energy. Considering the fact that zinc deficiency accelerates apoptosis and that excessive zinc accumulation inside cells results in a toxic effect that forces its death brings about several questions: Is intensification and acceleration of changes in zinc metabolism with age meaningful? Is there a real zinc deficiency occurring with age or in connection with the aforementioned pathological processes, or is it just a case of tissue and cell redistribution? When discussing factors that influence zinc homeostasis, can we consider zinc supplementation or regulation of zinc balance in the area of its redistribution? To clarify these aspects, an essential element will also be the clear understanding of the nomenclature used to describe changes in zinc balance. Zinc homeostasis can be different in different age groups and depends on sex, thus zinc dyshomeostasisrefers to changes in its metabolism that deviate from the normal rates for a particular age group and sex. This concept is very ample and implies that zinc deficiency may result from a low-zinc diet, poor absorption, excessive loss of zinc, zinc redistribution in intra- and extracellular compartments, or a combination of these factors that is inadequate for the given age and sex group. Such factor or factors need to be considered for preventing particular homeostasis disorders (or dyshomeostasis). Regulation of zinc metabolism by influencing reversal of redistribution processes ought to be the main point of pharmacologic and nonpharmacologic actions to reestablish zinc homeostasis. Supplementation and chelation are of marginal importance and can be used to correct long-term dietary zinc deficiency or zinc poisoning or in some cases in therapeutic interventions. In view of its biological importance, the problem posed by the influence of zinc metabolism requires further investigation. To date, one cannot consider, for example, routine zinc supplementation in old age, because changes of metabolism with age are not necessarily a cause of zinc deficiency. Supplementation is warranted only in cases in which deficiency has been established unambiguously. An essential element is to prevent sudden changes in zinc metabolism, which lead to dyshomeostasis in the terms defined here. The primary prophylaxes, regular physical activity, efficient treatment of chronic diseases, are all elements of such prevention.     

锌及锌转运蛋白ZnT3在小鼠海马苔藓纤维的一致性分布

目的 研究游离锌离子和锌转运蛋白ZnT3在小鼠海马的定位以及二的分布是否具有一致性。方法 应用锌TSQ荧光技术、锌金属自显影技术检测含锌神经元内的游离锌离子;应用免疫电镜技术检测ZnT3在含锌神经元轴突终末的分布。结果 游离锌离子和ZnT3免疫反应产物的分布在海马苔藓纤维内的分布具有一致性。在齿状回和CA3区的苔藓纤维内,锌和ZnT3蛋白定位于轴突终末的突触小泡。富含锌离子的含锌神经元轴突终末与CA3区锥体细胞的胞体和树突形成突触。尚可见锌离子存在于突触间隙内。结论 ZnT3向突触小泡内转运锌离子使锌离子聚积在含锌神经元轴突终末的突触小泡内,发挥锌离子的神经生物学功能。     

Carbon-dioxide exchange in lichens: determination of transport and carboxylation characteristics

Measurements were made of net rates of CO₂ assimilation in lichens at various ambient concentrations of CO₂ in air and in helox (79% He, 21% O₂). Because of the faster rate of CO₂ diffusion in the pores of lichen thalli when filled with helox than when filled with air, a given net rate of assimilation was achieved at a lower ambient concentration of CO₂ in helox. The differences were used to estimate resistances to diffusion through the gas-filled pore systems in lichens. The technique was first tested with five lichen species, and then applied in a detailed study with Ramalina maciformis, in which gas-phase resistances were determined in samples at four different states of hydration and with two irradiances. By assuming, on the basis of previous evidence, that the phycobiont in R. maciformis is fully turgid and photosynthetically competent at the smallest hydration imposed (equilibration with vapour at 97% relative humidity), and that, with this state of hydration, diffusion of CO₂ to the phycobiont takes place through continuously gas-filled pores, it was possible also to determine both the dependence of net rate of assimilation in the phycobiont on local concentration of CO₂ in the algal layer, and, with the wetter samples, the extents to which diffusion of CO₂ to the phycobiont was impeded by water films. In equilibrium with air of 97% relative humidity, the thallus water content being 0.5 g per g dry weight, the resistance to CO₂ diffusion through the thallus was about twice as large as the resistance to CO₂ uptake within the phycobiont. Total resistance to diffusion increased rapidly with increase in hydration. At a water content of 2 g per g it was about 50 times as great as the resistance to uptake within the phycobiont and more than two-thirds of it was attributable to impedance of transfer by water. The influences of water content on rate of assimilation at various irradiances are discussed. The analysis shows that the local CO₂ compensation concentration of the phycobiont in R. maciformis is close to zero, indicating that photorespiratory release of CO₂ does not take place in the alga, Trebouxia sp., under the conditions of these experiments.Symbols and Units rate of CO₂ diffusion in air relative to that in carrier gas (unity if the carrier gas is air and 0.43 if is helox) - A₁ net rate of CO₂ uptake by the lichen - A_p gross rate of carboxylation minus photorespiratory decarboxylation in the phycobiont, i.e. net rate of light-activated CO₂ exchange - A_* maximum, CO₂-saturated magnitude of A_p - c concentration of CO₂ - c_a ambient concentration of CO₂ - c_i c_a minus difference in CO₂ concentration across air-filled pore space in the thallus - c₈ CO₂ concentration equivalent to partial pressure of CO₂ at the surface of the phycobiont - ₁ magnitude of c_a at which A₁ = 0 - _* magnitude of c_* at which A_p = 0 - R rate of dark respiration in the lichen (mycobiont and phycobiont) - R rate of dark respiration in region between the surface of the lichen and an arbitrary distance from the surface within the thallus - r resistance to CO₂ transfer from lichen surface to the surface of the phycobiont - r resistance to CO₂ transfer between effective source of dark respiration in the lichen and the surface of the phycobiont - r_g, r _g components of r and r, respectively, attributable to transfer in air-phase - r_w, r _w components of r and r, respectively, attributable to transfer in water-phase - r component of r between surface of lichen and an arbitrary distance from the surface within the thallus - r_* resistance to CO₂ transfer and carboxylation in the phycobiont - RH relative humidity