特异切割12-脂加氧酶mRNA的核酶体外活性及动力学研究

根据锤头状核酶(Ribozyme)的作用模式,设计、合成并克隆了特异性切割12-脂加氧酶(12-LO)mRNA的核酶基因。以合成的25个核苷酸长的12-脂加氧酶RNA片段为底物与转录的核酶RNA一起保温检测其体外切割活性。实验结果表明,在37℃保温时,核酶在体外对12脂加氧酶具有较高的特异切割活性,其Km值为1300 nmol/L,其kcat值为0.083/min,在50℃保温时,核酶具有很高的切割活性,其kcat值为0.31/min。     

脱氧核酶的研究进展

具有RNA裂解活性的DNA分子称为脱氧核酶,它是经过体外选择技术经多次筛选获得的。脱氧核酶在切割与其互补的RNA底物分子时具有极高的特异性和切割效率,有望成为新的RNA灭活工具。     

脱氧核酶的研究进展

具有RNA裂解活性的DNA分子称为脱氧核酶。它是经过体外选择技术经多次筛选获得的。脱氧核酶在切割与其互补的RNA底物分子时具有极高的特异性和切割效率,有望成为新的RNA灭活工具。     

RNase P核酶对人巨细胞病毒UL54基因mRNA体外切割作用

外部引导序列(EGSs)是mRNA靶序列互补并引导RNaseP切割的小RNA片段。我们设计与人巨细胞病毒HCMV(Human Cytomegalovirus)UL54基因mRNA序列互补的EGSs,将其与大肠杆菌来源RNaseP催化核心M1RNA构建成M1GS核酶。通过对UL54基因亚克降片转录产物体外切割研究,证实该核酶具备对UL54 mRNA片段的特异切割能力,可以发展成为一种抗病毒试剂。     

Deoxyribozymes的研究进展

具有RNA裂解活性的DNA分子称为脱氧核酶,它是经过体外选择技术经多次筛选获得的。脱氧核酶在切割与其互补的RNA底物分子时具有极高的特异性和切割效率,有望成为新的RNA灭活工具。     

核酶在HIV—1基因治疗中的研究进展

HIV-1是一种特殊的逆转录病毒,其基因可转导入宿主细胞的基因内进行复制,核酶是一类具有催化活性的小分子RNA,能特异性与靶RNA分子结合后进行切割而抑制HIV-1,甚至一些RNA中间产物亦被核酶破坏。多项研究巳证实核酶可抑制细胞内HIV-1复制。与HIV-1的传统疗法相比,核酸基因治疗法干扰了病毒的装载和免疫系统的重建,能高效抑制病毒增殖,有助于患者免疫系统重建。     

RNase P核酶对人巨细胞病毒UL54基因mRNA体外切割作用

外部引导序列(EGSs)是mRNA靶序列互补并引导RNase P切割的小RNA片段.我们设计与人巨细胞病毒HCMV(Human Cytomegalovirus) UL54基因mRNA序列互补的EGSs,将其与大肠杆菌来源RNase P催化核心M1 RNA构建成M1GS核酶.通过对UL54基因亚克隆片转录产物体外切割研究,证实该核酶具备对UL54 mRNA片段的特异切割能力,可以发展成为一种抗病毒试剂.     

10-23脱氧核酶介导的生物传感器研究进展

近20年间,DNA介导的生物传感器得到了快速的发展,DNA能够作为遗传信息重要载体的同时,其折叠成的空间构象也具有很多的功能。功能核酸的概念逐渐引入到了包括生物传感、生物成像、医疗在内的重要领域中。10-23脱氧核酶作为功能核酸的一种,是通过体外筛选技术得到的,Mg2+存在的条件下能够特异性识别并切割RNA,切割位点为RNA中的嘧啶与嘌呤间的磷酸二酯键。由于其独特的识别以及切割能力,10-23脱氧核酶介导的相关疾病治疗得到了广泛的应用,同时人们逐渐开始关注10-23脱氧核酶介导的生物传感器的搭建。本文对于10-23脱氧核酶的结构、性质、作用方式以及改进修饰进行了介绍,并对10-23脱氧核酶介导的生物传感器的搭建以及应用进行了综述,为人们在未来使用10-23脱氧核酶搭建新型快捷生物传感器奠定理论基础。     

10-23脱氧核酶介导的生物传感器研究进展

近20年间,DNA介导的生物传感器得到了快速的发展,DNA能够作为遗传信息重要载体的同时,其折叠成的空间构象也具有很多的功能。功能核酸的概念逐渐引入到了包括生物传感、生物成像、医疗在内的重要领域中。10-23脱氧核酶作为功能核酸的一种,是通过体外筛选技术得到的,Mg²⁺存在的条件下能够特异性识别并切割RNA,切割位点为RNA中的嘧啶与嘌呤间的磷酸二酯键。由于其独特的识别以及切割能力,10-23脱氧核酶介导的相关疾病治疗得到了广泛的应用,同时人们逐渐开始关注10-23脱氧核酶介导的生物传感器的搭建。对于10-23脱氧核酶的结构、性质、作用方式及改进修饰进行了介绍,并对10-23脱氧核酶介导的生物传感器的搭建及应用进行了综述,旨为人们在未来使用10-23脱氧核酶搭建新型快捷生物传感器奠定理论基础。     

核酶对人巨细胞病毒mRNA片段的体外切割

引导序列GSs(GuideSequences)是能与mRNA互补,引导核酶RNaseP催化核心M1RNA对互补区域特异切割的小片段游离RNA。针对人巨细胞病毒HCMV(humancytomegalovirus)DNA聚合酶mRNA序列设计GS,共价结合到大肠杆菌来源M1RNA中,构建成M1GST7核酶。通过对巨细胞病毒DNA聚合酶亚克隆片段转录产物体外切割实验,表明该核酶具备对DNA聚合酶mRNA片段的特异切割能力 。     

INTERNAL PHLOEM IN THE PULVINUS OF SOYBEAN PLANTS

Glycine max, like many species of Fabaceae, has pulvini at the base of the petiole. In this structure, the vascular cylinder is constricted and consists of a ring of phloem surrounding a ring of xylem. A combination of light and transmission electron microscopy and histochemistry showed that, in addition, there are groups of internal phloem strands in the pulvinar pith. This was confirmed by direct observation of sieve plates and crystalline P-protein inclusions typical of leguminous sievetube members. Serial sections through the stem–pulvinus–petiole revealed that a spatial reorientation of the vascular tissue in the pulvinus resulted in the formation of internal phloem strands, which are continuous with the external phloem bundles above and below the pulvinus. Using 6(5)carboxyfluorescein (6CF) as a fluorescent tracer of phloem transport, we have shown that the internal phloem was active. In most of the experiments, when 6CF was applied to a source leaf, the internal phloem was not stained when the stem was girdled between the source leaf and the roots. Thus, we suggest that the internal phloem of the pulvinus of soybean is specialized for transport toward the root.     

EFFECTS OF AVIAN SEED DISPERSAL ON THE GENETIC STRUCTURE OF WHITEBARK PINE POPULATIONS

We used allozyme analysis to examine family structure, the spatial patterning of related individuals, in two populations of whitebark pine (Pinus albicaulis), a subalpine conifer that commonly displays a multistem form. The individual stems within clumps are genetically distinct individuals, having arisen from separate seeds. Individuals within a clump are genetically more similar than individuals in different clumps, but individuals in neighboring clumps do not appear to be more similar than individuals in distant clumps. This family structure appears to be a direct result of the seed-caching behavior of Clark's nutcrackers (Nucifraga columbiana), the primary dispersal agent for whitebark pine seeds.     

EVOLUTION OF MATE CHOICE FOR GENOME-WIDE HETEROZYGOSITY

The extent to which indirect genetic benefits can drive the evolution of directional mating preferences for more ornamented mates, and the mechanisms that maintain such preferences without depleting genetic variance, remain key questions in evolutionary ecology. We used an individual-based genetic model to examine whether a directional preference for mates with higher genome-wide heterozygosity ( H ), and consequently greater ornamentation, could evolve and be maintained in the absence of direct fitness benefits of mate choice. We specifically considered finite populations of varying size and spatial genetic structure, in which parent–offspring resemblance in heterozygosity could provide an indirect benefit of mate choice. A directional preference for heterozygous mates evolved under broad conditions, even given a substantial direct cost of mate choice, low mutation rate, and stochastic variation in the link between individual heterozygosity and ornamentation. Furthermore, genetic variance was retained under directional sexual selection. Preference evolution was strongest in smaller populations, but weaker in populations with greater internal genetic structure in which restricted dispersal increased local inbreeding among offspring of neighboring females that all preferentially mated with the same male. These results suggest that directional preferences for heterozygous or outbred mates could evolve and be maintained in finite populations in the absence of direct fitness benefits, suggesting a novel resolution to the lek paradox.     

缙云山大头茶幼苗种群构件结构及与环境因子的多元分析

基于构件理论,采用灰色关联度分析技术,对四川缙云山１９８９年风灾迹地林窗内大头茶（Ｇｏｒｄｏｎｉａａｃｕｍｅｎａｔａ）幼苗种群构件结构及其与环境因子的关系进行了研究。结果表明,四川缙云山大头茶幼苗种群构件结构主要分为一级枝、二级枝、当年生枝、空间结构、叶片等几大部分,分别可以以一级枝数或茎粗或长度、二级枝数或基粗或长度、当年生枝数、３年生一级枝数、总叶数等的变化特征来表达其动态特点。前四者间的相关性亦很高,后者（包括主茎上叶面积）和主茎上第一一级枝离地面高、主茎上第一叶距地高为比较稳定的特征,受其它生态因子作用影响不十分显著。相对而言,土壤全Ｎ、全Ｐ、全Ｋ、有机质含量及其ｐＨ值是比较关键的环境因子;而海拔高度和林自大小及地形坡度却比较次要。灰色关联度分析不失为一种比较简捷而有效的分析植物种群构件结构特征间及与环境因子间关系的方法。关键词     

THE INFLUENCE OF DENSITY AND SEX ON PATTERNS OF FINE-SCALE GENETIC STRUCTURE

Natal philopatry is expected to limit gene flow and give rise to fine-scale spatial genetic structure (SGS). The banner-tailed kangaroo rat ( Dipodomys spectabilis ) is unusual among mammals because both sexes are philopatric. This provides an opportunity to study patterns of local SGS faced by philopatric and dispersing animals. We evaluated SGS using spatial genetic autocorrelation in two D. spectabilis populations (Rucker and Portal) over a 14-year temporal series that covered low, medium, and high population densities. Significantly positive autocorrelation values exist up to 800 m at Rucker and 400 m at Portal. Density was negatively associated with SGS (low >medium >high), and suggests that increases in density are accompanied by greater spatial overlap of kin clusters. With regard to sex-bias, we find a small but significant increase in the SGS level of males over females, which matches the greater dispersal distances observed in females. We observed variation in SGS over the ecological time scale of this study, indicating genetic structure is temporally labile. Our study is the first temporal exploration of the influence of density and sex on spatial genetic autocorrelation in vertebrate populations. Because few organisms maintain discreet kin clusters, we predict that density will be negatively associated with SGS in other species.     

不同质地背景对自由飞行蜜蜂跟踪运动目标的影响

本文研究了不同质地背景对自由飞行蜜蜂跟踪运动目标的影响.质地背景采用了灰度调制度和空间频率均可调整的双正弦波图形构成,其灰度对比幅度为64级,平均灰度为32,运动目标是一个直径3.4厘米,灰度值为32的均匀灰色圆盘.当幅度调制度从小(31%)到大(93%)变化时,蜜蜂目标跟踪系统的水平增益S1也随之增大.这是由于较大的调制度可以给予蜜峰以较大的背景与目标之间的灰度对比.从短(7.6毫米)到长(38毫米)改变质地背景的空间波长,结果显示,在空间波长为30.4毫米处有一个明显的锐起峰值.计算结果表明,质地背景与目标之间在目标边缘的灰度反差,其最大值恰好发生在空间波长等于30.4毫米的情况下.这充分说明,S1的锐起峰值完全是由于形成于此的强烈灰度反差造成的.这一结果表明:无论目标运动与否,无论背景是均匀的还是质地的,背景与目标在目标边缘形成的灰度反差对蜜蜂发现并且跟踪目标起着重要乃至决定性的作用.     

荒漠绿洲过渡带植被空间异质性的可塑性面积单元问题

 以荒漠绿洲过渡带植被为研究对象,采用地统计理论与方法,以优势植物泡泡刺(Nitraria sphaerocarpa)和红砂(Reaumuria soongorica)种群为例,采用一个样地内2 500个10 m×10 m小样方中获取的植物盖度数据来研究采样尺度和划区方式对荒漠植被空间异质性的影响。其中尺度效应研究的思路是：一是保持粒度（样方）不变,增加幅度（样地）分析其对空间异质性的影响;二是保持幅度不变,改变粒度进行分析。划区效应是在样地尺度（500 m×500 m）上,把整块样地数据聚合成10 m×200 m、20 m×100 m、40 m×50 m、50 m×40 m、100 m×20 m和200 m×10 m等不同形状和不同方向的网格进行地统计分析。结果表明：荒漠绿洲过渡带植被空间异质性受采样尺度和划区方式影响显著。当样地面积小于200 m×200 m 时,所获取的数据不能真实反映泡泡刺种群的空间异质性;当样地面积小于100 m×100 m 时,所获取的数据不能真实反映红砂种群的空间异质性。调查泡泡刺和红砂种群的空间格局时,样方面积在20 m×20 m到30 m×30 m较为合适。植被空间异质性参数对划区方式的敏感性较强,其中泡泡刺种群的敏感性高于红砂种群。     

青藏高原东缘岷江冷杉天然群落的种群结构和空间分布格局

 岷江冷杉(Abies faxoniana)是青藏高原东缘亚高山顶极森林植被的优势种之一,主要分布于岷江、大渡河和白龙江的上游地区。该文研究了岷江冷杉天然原始群落的种群结构和空间分布格局。样方大小为100 m ×60 m。测定了所有个体的坐标及其胸径、高度和冠幅。将岷江冷杉按大小级分为5级,即幼苗：H（高度）<0.33 m; 幼树: H≥0.33 m, 且 DBH（胸径）<2.5 cm; 小树: 2.5 cm≤DBH<7.5 cm; 中树: 7.5 cm≤DBH <22.5 cm和大树: DBH≥22.5 cm。采用了Morisita 指数 (Iδ)、方差均值比 (V/m), 聚块度指标(m*/m)和空间点格局分析方法 (SPPA) (采用了Ripley二次分析法)4种方法分析岷江冷杉的空间分布格局。结果表明： 1) 岷江冷杉种群结构稳定。因为其年龄结构表现为增长型, 幼苗幼树储备丰富,密度分别为2 217·hm-2和2 683·hm-2,可见岷江冷杉天然更新良好,进而通过其“移动镶嵌循环”更新维持其种群的稳定性。在大小级结构图中的一些缺刻和年龄结构图中的“断代”现象,是干扰的时空异质性在采样的时间和空间断面上的反映。 2) 幼苗、幼树和小树在所有的研究尺度（从 1 m×1m 到 30 m×30 m）下都呈聚集分布。但中树和大树基本上呈随机分布。3) 聚集强度随尺度的变化而变化。上述的前3种方法表明,聚集强度随尺度的增加而减弱。但是,空间点格局分析法表明,岷江冷杉幼苗、幼树和小树的聚集强度首先随尺度的增加而增强,达到一定高峰后,随尺度的增加而减弱。 4) 岷江冷杉的空间分布格局是它与其自然环境长期作用的结果,同时也反映了其种群天然更新的格局和机制。 5) 4种分析方法对格局的判别基本一致, 但空间点格局分析法更能反映出格局强度随尺度的变化的关系,是值得推荐的一种分析空间分布格局的方法。使用空间点格局分析法的限制主要在于其计算和采样比较复杂。另外,由于Ripley 二次分析法对于“空白”的探测不敏感,需要进一步做一些方法上的改进。     

一种放大凋亡细胞Giemsa染色法及其在抗癌药物研究中的应用

本文介绍了一种用甲醇、冰醋酸、水处理细胞再经Giemsa染色的方法,经该方法处理后,细胞体积增大,凋亡细胞形态尤其胞核结构在普通光学显微镜下非常清晰.采用高三尖杉酯碱诱导四种不同类型肿瘤细胞凋亡,经放大细胞Giemsa染色之后,可获得清晰的不同时期凋亡细胞形态学表现及动态变化特征.进一步用琼脂糖电泳法检测四种细胞的DNA降解程度,结果表明,这种放大细胞的方法比琼脂糖电泳法更适用于形成凋亡小体之前细胞形态学特征的分析,为抗癌药物及细胞凋亡的研究提供了一种直观、准确的方法.     

Studies in Stomatal Behaviour: VI. AN INVESTIGATION OF THE LIGHT RESPONSES OF WHEAT STOMATA WITH THE ATTEMPTED ELIMINATION OF CONTROL BY THE MESOPHYLL

As in the previous investigation (Part I), stomatal responsesin wheat were followed by means of porometer readings; betweenthese, air was forced through the stomata to sweep out the substomatalcavities. Instead of forcing the whole of a relatively slowair-flow in through the stomata, however, a fast flow was sweptacross the leaf surface and a portion of this passed into theintercellular space system. This helped to maintain the desiredcarbon dioxide content close to the stomatal openings and thestomata could thus be treated with different carbon dioxideconcentrations. A factorial experiment was carried out in which6 carbon dioxide concentrations from zero to 0084 per cent,were combined with 3 light intensities from 90 to 800 f.c, the18 treatments being replicated 6 times in three ‘66 latinsquares’. A supplementary experiment in which the same6 carbon dioxide treatments were given in darkness was carriedout in duplicate. The results confirmed previous findings inthat the stomata showed no appreciable response to variationof external carbon dioxide supply between zero and 001 percent, concentration and that a large and highly significantlight response was found at each of these two carbon dioxidelevels. This is held to indicate the existence of a light effect(direct or indirect) not operating by reduction of the internalcarbon dioxide content of the guard cells. On the assumptionthat the rates of air flow through the stomata were adequate,the combined results of the two experiments are interpretedmainly in terms of two direct effects of light upon the stomata,one depending on photosynthetic reduction of carbon dioxidein the guard cells and the other, mentioned above, not so operating.To these must be added the indirect effect responsible for the‘transmission’ described in Part 1, which mightpossibly account for the whole of the second direct effect postulated.Finally, if the rates of flow were in fact grossly inadequate,the trends attributed to the first direct effect mentioned abovemight have been due to indirect control by the mesophyll viathe intercellular space atmosphere. Such control must accountfor part of the light responses which occur under natural conditions.The natural light response is thus made up of at least two andmore probably four components—two direct and two indirect,in each case one operating by the internal carbon dioxide andthe other not. A highly significant diurnal rhythm of stomatalmovement under constant illumination and temperature is shownto occur in wheat. The measure of stomatal resistance used (logresistance per 10,000 stomata) is discussed, and it is concludedthat although it is empirical a much more satisfactory measureis not at present available.