紫外线B区对小牛胸腺DNA损伤的拉曼光谱研究

用拉曼光谱检测了远紫外区UVB(280m～320nm)对小牛胸腺DNA的损伤,并对这个区段紫外线对DNA的不同照射时间时的损伤特征进行了比较分析。实验中所用的紫外线强度与太阳光强度相当。结果表明,辐照3h以内时,UVB对DNA的构型有较明显的损伤,这可能是受到嘧啶碱基损伤的影响。相对而言,UVB对脱氧核糖和碱基的损伤要严厉得多。就碱基对的受损伤程度来说,嘧啶碱受损最严重,部分证明了环丁烷嘧啶二聚体和6-4光产物的形成。经过3h UVB照射,AT碱基对和和胞嘧啶环的堆叠程度有所瓦解,一些碱基对受到修饰。而一些拉曼特征峰强度的反复波动,则说明了长时间的紫外照射可以导致部分DNA光复活的产生。进一步分析发现,UVB以一种较快的方式对DNA产生损伤。     

5-~(131)碘尿嘧啶在细菌脱氧核糖核酸复制研究中的应用

本文报导了一个用5-~(131)碘尿嘧啶代替[~3H]-胸腺嘧啶进行细菌DNA复制研究的新方法。通过对放射性参入产物的碱(KOH)水解和酶(DNase和RNase)水解的实验证明:5-~(131)碘尿嘧啶与[~3H]-胸腺嘧啶一样能特异地参入大肠杆菌胸腺嘧啶缺陷变异株的DNA,而不能参入其RNA。对氨基酸饥饿同步化的大肠杆菌15T~-,当应用5-~(131)碘尿嘧啶的参入来测定复加氨基酸后的DNA复制起步时间时,获得的结果与使用[~3H]-胸腺嘧啶脱氧核苷参入的结果相一致。天然的DNA碱基成分胸腺嘧啶强烈地竞争性地抑制5-~(131)碘尿嘧啶的参入能力,而天然的RNA碱基成分尿嘧啶则影响很小。此外,5-碘尿嘧啶对大肠杆菌胸腺嘧啶缺陷变异株的生长有抑制作用,但这种抑制要在加入5-碘尿嘧啶后2小时才明显地产生,而5-碘尿嘧啶对大肠杆菌野生株的生长没有影响。实验结果表明:在细菌DNA复制的研究中,5-~(131)碘尿嘧啶参入的方法与使用[~3H]-胸腺嘧啶参入的方法有同样的可靠性,其优点是由于~(131)碘辐射γ射线,故放射性参入样品的制备和测定都比较简便,因而,它比[~3H]-胸腺嘧啶更适合于有关临床和实验室的使用。     

阳光紫外辐射对两种微藻类光化学效率的影响

近年来人类活动释放的含氟、氯等有机物对臭氧层造成破坏,使得到达地球表面的紫外线B（Ultra-violet B（UV—B）．280-315nm）辐射增强。为此,生物活动受到了多方面的影响。以往的研究发现,紫外辐射（Ultra-violet radiation．UVR）可以抑制藻类的生长及光合固碳和游动性（有鞭毛的种类）。并损伤细胞色素和遗传物质DNA。     

细菌的GC%鉴定法

<正>DNA的化学组成及构造 脱氧核糖核酸(DNA)担负着生物的所有遗传信息,从细胞水平来看,它可能是信息表达的兰图,细胞分裂时形成两个相同的分子(这叫做复制),分配给两个子细胞,使之遗传下去。其化学基本单位是由1分子脱氧核糖,1分子磷酸及1分子碱基(4种中的某一种)所构成,称为核苷酸。如图1所示,4种碱基乃是2种嘌呤碱基,即腺嘌呤、鸟嘌呤和2种嘧啶碱基,即胸腺嘧啶,胞嘧啶。腺嘌呤与胸腺嘧啶之间能形成     

2005年高考生物学备考试题--"遗传与进化"部分

一、选择题:(本题包括28小题,每题1.5分,共42分。每小题只有一个选项最符合题意)1.孟德尔做了如图所示的杂交实验,决定图中种子种皮基因型的是()。A.由植株A和植株B共同决定B.只由植株A决定C.只由植株B决定D.与植株A或植株B无关2.与“阿波罗登月计划”相提并论的“人类基因组计划”的主要任务是测定人体基因组整体序列。决定基因遗传特异性的是()。A.基因的碱基对排列顺序B.嘌呤总数与嘧啶总数的比值C.碱基互补配对的原则D.脱氧核苷酸链上磷酸和脱氧核糖的排列特点3.理论上同种生物同一个体每个表皮细胞与神经细胞内所含DNA是相同的,…     

1991年普通高等学校招生全国统一考试生物学试题

一、选择题:本大题共35个小题,每小题1分,共35分。在每小题给出的四个选项中,只有一项是符合题目要求的。把所选项前的字母填在括号内。 1.人的皮肤痛觉感受器位于( ) A.角质层 B.生发层 C.真皮 D.皮下组织 2.下列哪一组物质是DNA的组成成分?( ) A.脱氧核糖、核酸和磷酸 B.脱氧核糖、碱基和磷酸 C.核糖、碱基和磷酸 D.核糖、嘧啶、嘌呤和磷酸 3.在人体血液循环系统中,下列哪条血管的血液中     

增强UV-B辐射与干旱复合处理对小麦幼苗生理特性的影响

为研究由于平流层臭氧层减薄紫外线B辐射增强在干旱地区对春小麦生理特性的影响的特殊性,模拟平流层臭氧减少20％时辐射到地表的紫外线B（UV－B,280－315nm)的增强和水分胁迫（－0.5Mpa,聚乙二醇PEG－6000处理获得）,通过测定两种胁迫下春小麦（Triticum aestivum L.)叶绿素含量、类黄酮含量、水势、细胞膜相对透性、超氧歧化酶（SOD）活性及丙二醛（MDA）含量,研究了增强UV－B辐射和水分胁迫复合作用对温室种植的小麦幼苗生理生化的影响。实验结果表明,虽然水分胁迫和UV－B辐射单独或复合处理都使春小麦的叶绿素含量降低,但仅UV－B辐射增强单独处理显著地降低小麦叶绿素a、b和总叶绿素的含量,而水分胁迫以及复合处理对叶绿素的含量的降低作用不显著。两种胁迫无论是单独作用还是复合作用均能使类黄酮含量升高,并且处理第3天比第1天高出近50％,复合处理下类黄酮的含量大于两个因子单独处理。UV－B辐射和水分胁迫处理1d对春小麦叶片的相对电导率的影响不明显,处理3d后两种胁迫下相对电导率均上升,表明膜透性增加,其中水分胁迫作用下增加尤其明显。膜质过氧化产物丙二醛的含量,在两种因子单独和复合作用下都升高,说明膜的生理功能受到了一定的不利影响。活性氧清除剂超氧化物歧化酶（SOD）的活性在各种处理下,都没有发生改变。虽然增强的UV－B辐射和干旱处理一样,会显著降低植物水势,但是,UV－B辐射与干旱同时处理时叶片水势降低的程度,不但没有比两者分别处理时降低程度之和低,而且比单做干旱处理时的降低程度还低,这表明UV－B辐射和干旱胁迫同时处理时,UV－B辐射不但没有加重水分胁迫,反而减轻了干旱对春小麦生长的胁迫。由此可以认为,在干旱条件下,增强UV－B辐射不会加剧而是有利于提高小麦对干旱的抗性。     

T4核酸内切酶V的分离纯化

 <正> T_4核酸内切酶V[endodeoxyribonuclease(Pyrimidine dimer)EC 3.1.25.1,简称Endo V]是由噬菌体T_4感染宿主E.coli后产生的一种修复酶。它能特异识别紫外线(UV)辐射在DNA引起的损伤产物环丁烷嘧啶二聚体(PD),在组成PD的两个嘧啶核苷酸之间切割磷酸二酯键,造成单链缺刻,从而启动E.coli体内的切除修复途径~[1]。Endo V不仅具有对PD的高     

UV-B辐射和蒽对三角褐指藻DNA伤害的相互作用

运用生态毒理学和生物化学的方法研究了紫外线和多环芳烃－蒽对三角褐指藻DNA的伤害作用,结果表明,蒽对三角褐指藻的生长有抑制作用;随着蒽浓度的增加,三角揭褐藻DNA损伤程度增加;在蒽浓度固定不变时,随着处理时间的延长,DNA的损伤程度同样提高;在蒽的处理过程中同时伴有紫外线的辐射处理,DNA的损伤程度加剧;蒽处理解除一段时间后,DNA损伤程度未明显减轻、而UV－B处理解除后,DNA的损伤可明显恢复,说明DNA的损伤可在一定程度上指示海洋微藻受蒽伤的程度。     

UV-B辐射对植物花粉萌发率和花粉管生长的累积效应

研究了19种植物花粉在不同UV－B辐射强度和辐照时间下其萌发率和花粉管伸长的变化,结果表明,UV－B辐射增加显著抑制大多数植物花粉的萌发率和花粉管生长;与对照相比,较高强度的UV－B对花粉的抑制作用大于较低强度;几个种的花粉萌发率及花粉管生长对UV－B增强不敏感,甚至被UV－B辐射所促进;辐射时间越长,对花粉抑制作用愈大,说明具有辐射累积效应,由此可知,植物花粉的萌发过程对UV－B的敏感性变化在自然条件下将会产生严重的生态学后果。     

Artificial and solar UV radiation induces strand breaks and cyclobutane pyrimidine dimers in Bacillus subtilis spore DNA

The loss of stratospheric ozone and the accompanying increase in solar UV flux have led to concerns regarding decreases in global microbial productivity. Central to understanding this process is determining the types and amounts of DNA damage in microbes caused by solar UV irradiation. While UV irradiation of dormant Bacillus subtilis endospores results mainly in formation of the "spore photoproduct" 5-thyminyl-5,6-dihydrothymine, genetic evidence indicates that an additional DNA photoproduct(s) may be formed in spores exposed to solar UV-B and UV-A radiation (Y. Xue and W. L. Nicholson, Appl. Environ. Microbiol. 62:2221-2227, 1996). We examined the occurrence of double-strand breaks, single-strand breaks, cyclobutane pyrimidine dimers, and apurinic-apyrimidinic sites in spore DNA under several UV irradiation conditions by using enzymatic probes and neutral or alkaline agarose gel electrophoresis. DNA from spores irradiated with artificial 254-nm UV-C radiation accumulated single-strand breaks, double-strand breaks, and cyclobutane pyrimidine dimers, while DNA from spores exposed to artificial UV-B radiation (wavelengths, 290 to 310 nm) accumulated only cyclobutane pyrimidine dimers. DNA from spores exposed to full-spectrum sunlight (UV-B and UV-A radiation) accumulated single-strand breaks, double-strand breaks, and cyclobutane pyrimidine dimers, whereas DNA from spores exposed to sunlight from which the UV-B component had been removed with a filter ("UV-A sunlight") accumulated only single-strand breaks and double-strand breaks. Apurinic-apyrimidinic sites were not detected in spore DNA under any of the irradiation conditions used. Our data indicate that there is a complex spectrum of UV photoproducts in DNA of bacterial spores exposed to solar UV irradiation in the environment.     

Artificial and Solar UV Radiation Induces Strand Breaks and Cyclobutane Pyrimidine Dimers in Bacillus subtilis Spore DNA

The loss of stratospheric ozone and the accompanying increase in solar UV flux have led to concerns regarding decreases in global microbial productivity. Central to understanding this process is determining the types and amounts of DNA damage in microbes caused by solar UV irradiation. While UV irradiation of dormant Bacillus subtilis endospores results mainly in formation of the “spore photoproduct” 5-thyminyl-5,6-dihydrothymine, genetic evidence indicates that an additional DNA photoproduct(s) may be formed in spores exposed to solar UV-B and UV-A radiation (Y. Xue and W. L. Nicholson, Appl. Environ. Microbiol. 62:2221–2227, 1996). We examined the occurrence of double-strand breaks, single-strand breaks, cyclobutane pyrimidine dimers, and apurinic-apyrimidinic sites in spore DNA under several UV irradiation conditions by using enzymatic probes and neutral or alkaline agarose gel electrophoresis. DNA from spores irradiated with artificial 254-nm UV-C radiation accumulated single-strand breaks, double-strand breaks, and cyclobutane pyrimidine dimers, while DNA from spores exposed to artificial UV-B radiation (wavelengths, 290 to 310 nm) accumulated only cyclobutane pyrimidine dimers. DNA from spores exposed to full-spectrum sunlight (UV-B and UV-A radiation) accumulated single-strand breaks, double-strand breaks, and cyclobutane pyrimidine dimers, whereas DNA from spores exposed to sunlight from which the UV-B component had been removed with a filter (“UV-A sunlight”) accumulated only single-strand breaks and double-strand breaks. Apurinic-apyrimidinic sites were not detected in spore DNA under any of the irradiation conditions used. Our data indicate that there is a complex spectrum of UV photoproducts in DNA of bacterial spores exposed to solar UV irradiation in the environment.     

紫外辐射诱导植物叶片DNA损伤敏感性差异

单细胞凝胶电泳(彗星检测, comet assay)技术已广泛应用于动物细胞DNA损伤检测, 但在植物细胞DNA损伤检测中的应用尚不多见。本研究通过对动物细胞彗星检测方法的改进, 利用植物细胞原生质体作为材料, 研究了不同发育期九里香(Murraya panicuata)叶片对UV-B诱导的DNA损伤的敏感性差异。彗星检测结果表明, 九里香叶片DNA的损伤程度与UV-B辐射的剂量呈正相关; 在相同UV-B辐射剂量下, 九里香幼嫩叶片比成熟叶片的DNA损伤量大, 表明其幼嫩叶片对UV-B辐射的敏感性比成熟叶片高。     

紫外辐射诱导植物叶片DNA损伤敏感性差异

单细胞凝胶电泳（彗星检测,cometassay）技术已广泛应用于动物细胞DNA损伤检测,但在植物细胞DNA损伤检测中的应用尚不多见。本研究通过对动物细胞彗星检测方法的改进,利用植物细胞原生质体作为材料,研究了不同发育期九里香（Murraya panicuata）叶片对UV-B诱导的DNA损伤的敏感性差异。彗星检测结果表明,九里香叶片DNA的损伤程度与UV-B辐射的剂量呈正相关：在相同UV—B辐射剂量下,九里香幼嫩叶片比成熟叶片的DNA损伤量大,表明其幼嫩叶片对UV-B辐射的敏感性比成熟叶片高。     

The two major spore DNA repair pathways, nucleotide excision repair and spore photoproduct lyase, are sufficient for the resistance of Bacillus subtilis spores to artificial UV-C and UV-B but not to solar radiation.

Bacterial endospores are 1 to 2 orders of magnitude more resistant to 254-nm UV (UV-C) radiation than are exponentially growing cells of the same strain. This high UV resistance is due to two related phenomena: (i) DNA of dormant spores irradiated with 254-nm UV accumulates mainly a unique thymine dimer called the spore photoproduct (SP), and (ii) SP is corrected during spore germination by two major DNA repair pathways, nucleotide excision repair (NER) and an SP-specific enzyme called SP lyase. To date, it has been assumed that these two factors also account for resistance of bacterial spores to solar UV in the environment, despite the fact that sunlight at the Earth's surface consists of UV-B, UV-A, visible, and infrared wavelengths of approximately 290 nm and longer. To test this assumption, isogenic strains of Bacillus subtilis lacking either the NER or SP lyase DNA repair pathway were assayed for their relative resistance to radiation at a number of UV wavelengths, including UV-C (254 nm), UV-B (290 to 320 nm), full-spectrum sunlight, and sunlight from which the UV-B portion had been removed. For purposes of direct comparison, spore UV resistance levels were determined with respect to a calibrated biological dosimeter consisting of a mixture of wild-type spores and spores lacking both DNA repair systems. It was observed that the relative contributions of the two pathways to spore UV resistance change depending on the UV wavelengths used in a manner suggesting that spores irradiated with light at environmentally relevant UV wavelengths may accumulate significant amounts of one or more DNA photoproducts in addition to SP. Furthermore, it was noted that upon exposure to increasing wavelengths, wild-type spores decreased in their UV resistance from 33-fold (UV-C) to 12-fold (UV-B plus UV-A sunlight) to 6-fold (UV-A sunlight alone) more resistant than mutants lacking both DNA repair systems, suggesting that at increasing solar UV wavelengths, spores are inactivated either by DNA damage not reparable by the NER or SP lyase system, damage caused to photosensitive molecules other than DNA, or both.     

Marine Bacterial Isolates Display Diverse Responses to UV-B Radiation

The molecular and biological consequences of UV-B radiation were investigated by studying five species of marine bacteria and one enteric bacterium. Laboratory cultures were exposed to an artificial UV-B source and subjected to various post-UV irradiation treatments. Significant differences in survival subsequent to UV-B radiation were observed among the isolates, as measured by culturable counts. UV-B-induced DNA photodamage was investigated by using a highly specific radioimmunoassay to measure cyclobutane pyrimidine dimers (CPDs). The CPDs determined following UV-B exposure were comparable for all of the organisms except Sphingomonas sp. strain RB2256, a facultatively oligotrophic ultramicrobacterium. This organism exhibited little DNA damage and a high level of UV-B resistance. Physiological conditioning by growth phase and starvation did not change the UV-B sensitivity of marine bacteria. The rates of photoreactivation following exposure to UV-B were investigated by using different light sources (UV-A and cool white light). The rates of photoreactivation were greatest during UV-A exposure, although diverse responses were observed. The differences in sensitivity to UV-B radiation between strains were reduced after photoreactivation. The survival and CPD data obtained for Vibrio natriegens when we used two UV-B exposure periods interrupted by a repair period (photoreactivation plus dark repair) suggested that photoadaptation could occur. Our results revealed that there are wide variations in marine bacteria in their responses to UV radiation and subsequent repair strategies, suggesting that UV-B radiation may affect the microbial community structure in surface water.     

Application of the comet assay to measure DNA damage induced by UV radiation in the hydrophyte, Spirodela polyrhiza

The single-cell gel electrophoresis or comet assay is now widely used to detect DNA damage in animal cells induced by radiation or chemicals. Here, we apply the comet assay to measure ultraviolet (UV)-B-induced DNA damage in plant cells. The accepted animal cell protocol for the comet assay was modified to adapt it to plant cells. The major modifications were conversion of the plant cells to protoplasts and the use of T4 endonuclease V. As a positive control hydrogen peroxide was applied. Significant DNA damage was detected at 100 μ M H₂O₂. This type of DNA damage was not affected by T4 endonuclease V treatment, which implies that the mechanism of H₂O₂-induced DNA damage was different from UV-B-induced DNA damage. Our results also indicate that both UV-A and UV-B radiation can induce DNA single-strand breaks in plant cells, while UV-B was more effective than UV-A for inducing pyrimidine dimer formation.     

UV damage to plant life in a photobiologically dynamic environment: the case of marine phytoplankton

The effect of UV-B radiation on growth of marine phytoplankton was investigated in relation to DNA damage induced by a range of biologically effective doses (BEDs). Emiliania huxleyi (Prymnesiophyceae) was chosen as a model organism of the ocean's phytoplankton because of its importance in global biogeochemical cycling of carbon and sulphur, elements that influence the world's climate as components of the trace gases carbon dioxide (CO₂) and dimethylsulfide (DMS). A marine diatom, Cyclotella, was studied for its capacity to repair the DNA damage, quantified as thymine dimers by the application of a monoclonal antibody against these photoproducts. DNA repair was shown to be complete after just a few hours of exposure to visible light; the repair rate increased with PAR intensity. E. huxleyi appeared to be most sensitive to UV-B radiation: growth was already affected above a dose of 100 J m^-2 d^-1 (biologically effective radiation, weighted with Setlow's DNA action spectrum), probably through effects on the cell cycle related to damage to nuclear DNA: mean specific growth rates were inversely correlated with thymine dimer contents in cells. Near the ocean's surface UV-B radiation conditions that induce the changes observed by us in cultures can be expected during the growing season of phytoplankton, not only in the tropics but also at higher latitudes. Nevertheles, blooms of species such as E. huxleyi are often excessive in the field. It is suggested that exposure duration of cells near the surface of the ocean can be shorter than our artificial 3 h in the laboratory due to vertical mixing, a phenomenon that is typical for the ocean's upper 50–100 m. When mixing reaches depths greater than the layer where most UV-B is attenuated, negative effects on cells through UV-A-induced inhibition of photosynthesis may prevail over DNA damage, the action spectrum of which has been shown to be limited to the UV-B part of the spectrum. Moreover, the radiation wavelengths that induce DNA damage repair (UV-A and visible) are attenuated vertically much less than UV-B. The photobiological situation in the upper ocean is much more complicated than on land, and effects of UV radiation on plankton biota can only be modelled realistically here when both the spectrally differential attenuation in the UV and visual part of the spectrum and the rate of vertical mixing are taken into account. Action spectra of both damage and repair of DNA and of photosynthesis inhibition of representative microalgal species are the second conditio sine qua non if we want to predict the effect of stratospheric ozone depletion on marine phytoplankton performance.     

Specificity and Photomorphogenic Nature of Ultraviolet-B-Induced Cotyledon Curling in Brassica napus L

Three general classes of photomorphogenic photoreceptors have been characterized in higher plants: phytochrome, a blue light/ultraviolet (UV)-A photoreceptor(s), and a UV-B sensory system(s). Although a great deal is known about phytochrome and the blue light/UV-A photoreceptor(s), little is known about UV-B detection processes. One reason for this is the lack of readily quantifiable morphogenic responses that are specifically induced by UV-B radiation. We have discovered a response to UV-B, upward curling of Brassica napus L. cotyledons, that may be useful for probing the mechanism of UV-B photoreception. The process was initially observed when B. napus seeds were germinated under visible light plus UV-B radiation, but did not occur under visible light alone or visible light plus UV-A. When 5-d-old seedlings grown in visible light were given relatively short exposures of UV-B (100 min of 5.5 [mu]mol m-2 s-1), the curling response was also observed. Development of curling was separated from the application of this UV-B pulse by a 14-h latent period. Pulses of red light, blue light, farred light, and UV-A (100 min of 5.5 [mu]mol m-2 s-1) did not induce curling, indicating UV-B specificity Additionally, these other spectral regions did not reverse or enhance the UV-B-triggered response. The degree of curling showed a log-linear dependence on UV-B fluence (6-40 mmol m-2) and reciprocity with respect to length of exposure and fluence rate. The data indicate that curling is photomorphogenic in nature and may be triggered by a single photoreceptor species.