用地高辛标记Y染色体特异探针检测人的性别

采用了一种新的DNA探针非同位素标记方法,即地高辛配基标记人Y染色体特异DNA探针,成功地用于人基因组中男性特异DNA的检测。同时,将此种方法与生物素标记探针方法作了比较。结果表明：地高辛配基标记探针方法优于生物素标记。     

分子生物学技术讲座（五）：放射性标记的DNA和RNA探针的制备

放射性标记的核酸探针（DNA和RNA探针）目前已被广泛地应用于分子生物学的各个领域,例如克隆的筛选,Southern杂交分析,基因表达水平的测定等等。放射性核酸分子探针是指特定的已知核酸分子片段,内含放射性核素（例：‘千、‘H和”S）,并能与被检测的核酸分子退火杂交（核酸序列互补）,因此可用于待测核酸样品中特定基因序列片段的探测。1探针的种类和选择根据核酸分子探针的来源及其性质可将其分成3大类,即：DNA探针、RNA探针和人工合成的寡聚核青酸探针。而DNA探针又可分为基因组DNA探针和。DNA探针。探针的选择是根据不…     

RecA蛋白介导的三链核酸结构形成及其序列提取

人工合成的单链DNA分子经PCR扩增形成双链DNA分子。将RecA蛋白与生物素标记的寡聚核酸探针序列在ATPγS存在的情况下共同哺育,使RecA蛋白包裹寡聚核酸探针,然后加入含同源序列的上述双链DNA分子经适当环境哺育形成了稳定的局部三链核酸结构。通过加入链亲和素包裹的磁珠吸附生物素化的探针,这样同源双链DNA分子与寡聚核酸探针形成的局部三链核酸结构也被吸附在磁珠上。使用磁分离装置提取这一结构,逐步降低盐离子浓度以洗脱双链DNA分子。将洗脱液中残留的蛋白质去除,经PCR扩增可获得目的DNA序列。同时使用同源探针和非同源探针在其它序列中提取目的DNA序列,结果显示目的DNA序列只被同源探针提取。实验结果显示了这一三链核酸结构形成的序列特异性,并且其稳定性随盐离子浓度降低而下降。提示在这一结构中同源的寡聚核酸单链与双链DNA分子形成了氢键结合,同时提示使用文中描述的方法可以提取特异的序列,用以克隆相应的基因。     

地高辛标记DNA探针杂交法检测Vero细胞DNA残留量的适用性验证及应用

目的对地高辛标记DNA探针杂交法检测人用狂犬病疫苗(Vero细胞)DNA残留量进行适用性验证及应用。方法对地高辛标记DNA探针杂交法检测人用狂犬病疫苗(Vero细胞)DNA残留量进行特异性、灵敏度及稳定性验证,并应用该方法检测3批人用狂犬病疫苗(Vero细胞)的DNA残留量。结果地高辛标记探针的标记效率为0.1 pg。验证结果显示探针与非同源DNA无杂交;最低检测限度为1 pg;探针在-20℃放置7个月后,检测灵敏度仍可达到1 pg;3批人用狂犬病疫苗(Vero细胞)中残留DNA含量均符合《中国药典》规定质量控制标准。结论地高辛标记DNA探针杂交法特异性、灵敏度好,结果稳定,适用于人用狂犬病疫苗(Vero细胞)中Vero细胞DNA残留量的检测及疫苗生产过程和其成品的质量控制,对其他以Vero细胞为基质的病毒性疫苗质量控制具有借鉴意义。     

生物素标记DNA探针杂交条件探讨

DNA探针技术正越来越广泛地应用于医学基础研究和临床检测.生物素标记探针已在某些领域部分取代同位素标记探针而发挥作用。杂交是DNA探针技术中重要一环.各种杂交条件的变化均会对杂交结果并最终对检测结果产生影响。文献中关于杂交条     

卫生工业厂商协会(HIMA)DNA杂种探针会议要点

最近召开的名为“诊断试验的尖端:DNA探针”的专题会议,回顾了DNA杂交种探针的现状和将来的趋势。本文是卫生工业厂商协会医学科学部生物技术委员会发起的于1984年6月21日到22日在弗吉尼亚州的阿林顿召开的会议要点。在回顾DNA探针的基本技术时,Enzo Biochem公司研究部主任Dean Engelhardt提到,在双相测定中,杂交速率比靶DNA和探针都在溶液里慢2～10倍。双相测定中的靶DNA如同现在市售的探针一样,被固定在固体支持物上。所以,在做DNA杂交时,人们希望采用较现行双相法为快的单相测定法。在标记DNA探针方面,Enzo正在研制能将信息分子附     

综合遗传学公司推出食品沙门氏菌快速诊断试验盒

综合遗传学(Integrated Genetics)公司最近推出它的第一个产品--检出食品沙门氏菌用的快速诊断试验盒。它被称为GENE-TRAK,是以DNA杂交技术为基础的。目前只能用于沙门氏菌的检出。该公司的科学家已分离出DNA探针,含有与沙门氏菌DNA互补的碱基序列模式。分离出可能存在于食品样品中的任何微生物的DNA链后,将沙门氏菌DNA探针加入样品中,若探针与存在于样品中的DNA单链杂交,则表明其中有沙门氏菌存在。     

酶标HBV DNA探针的研制与应用

本实验采用一种非放射性物质——碱性磷酸酶标记乙肝病毒HBV DNA制备分子探针。碱性磷酸酶在苯醌作用下与单链DNA联结,形成DNA和酶的共价复合物,即酶标探针。此探针通过分子杂交与待测DNA结合,与酶的底物作用显色,几小时内可观察结果,其最低检测量约为10pg。用此探针检测乙肝病人血清中的HBV DNA,与~(32)P标记的探针比较,酶标探针可检测出~(32)P标记探针检出率的95.7％。结果表明,所合成的酶标探针具有准确、简便、快速、安全而经济的优点,具有应用前景。     

人巨细胞病毒的生物素DNA探针的制备和应用

本研究用克隆的HCMV AD169株DNA片段,制备了生物素标记的DNA探针,建立了检测临床脐带血、尿标本中HCMV DNA的核酸探针杂交方法。该探针可测出100pg同源DNA,不与人胚肺细胞、Hep-2细胞DNA以及其他疱疹病毒的DNA发生反应。用核酸杂交方法检测了30份脐带血标本,有11例阳性,阳性率为33％。10例孕妇尿标本中,3例阳性,阳性率为30％。检测结果表明：我们建立的生物素标记的HCMV DNA探针的点杂交法,具有高度的特异性、敏感性,比分离病毒法更迅速,可用于HCMV感染的临床标本的病毒核酸检测。     

烟草花叶病毒辽宁分离物Northern杂交检测体系的建立

为了建立烟草花叶病毒辽宁分离物(TMV-LN)的高特异性、高灵敏度的分子杂交检测体系,从粗提纯的TMV-LN粒子中提取RNA,设计特异性引物通过RT-PCR扩增TMV-LN的CP和3'端非翻译区域,将片段连至p UC119载体获得重组质粒p UCTMV-PP,体外转录获得地高辛(DIG)标记的TMV-LN正义链RNA杂交检测探针,同时构建DNA检测探针作为对照。采用点印迹(Dot-blot)杂交和Northern杂交对比RNA探针和DNA探针对TMV-LN的检测特异性和灵敏性。检测结果表明,RNA探针和DNA探针在点印记杂交和Northern杂交中均表现出良好的检测特异性,RNA探针在检测灵敏度方面要略好于DNA探针,且点印迹杂交体系在病毒定性方面较为快捷,Northern杂交体系在病毒基因组RNA的定量方面具有明显优势。     

Systematic Study on Lomatogonium A. Br. (Gentianaceae)

The present paper deals with the infrageneric classification, phylogeny and geographic distribution of the genus Lomatogonium. A cladistic analysis was undertaken to establish the taxa and to evaluate the relationships between the taxa. The PAUP computer program was used in this analysis. The most parsimonious tree (Cladogram) of the rotate-corolla group of subtribe Gentianinae shows that Lomatogonium is closely related to Lomatogoniopsis and Swertia, but distantly to Veratrilla. Among them, Swertia is more primitive than Lomatogonium and hence Sect. Swertia was selected as the outgroup to polarize the character states of ingroup (Lomatogonium). A data matrix of 29 charaters of Lomatogonium was made for constructing the cladogram. Two most parsimonious trees were formed one of which, with the lowest f value, was at last selected as a shortest tree. In this tree 18 species fall into three groups, i.e. Sect. Sarcorhizoma, Sect. Lomatogonium and Sect. Pleurogynella. The former comes at a lower level with more plesiomorphies while the latter at a higher level with more apomorphies. Lomatogonium is distributed in the northern temperate zone. However, 16 species are centred in Asia and two extend to Europe, or further to the Arctic region, but none has been found from Africa, Australia and South erica. The analysis of distribution pattern of species shows that the Qinling-Hengduan Mountain region is both the frequence and diversity centers of Lomatogonium. From the cladogram of Lomatogonium (Fig. 5 ), L. perenne appears to occupy the most plesiomorphic node. This is an indication that it is the extant species closest to the ancestral form and it also implies that the ancestral species may reside in the habitat of this species (the Qinlin-Hengduan Mountain region). On the other hand, a umber of species of Swertia Sect. Swertia also occur in this region today, which indicates that the Qinlin-Hengduan Mountain region may well be the original center of Lomatogonium. From the distribution pattern of L. rotatum, it can be concluded that the time of the origin dates back at least before the Pliocene. After emergence, this genus had first developed and dispersed in the original center and adjacent region, then diverged into two lineages. One gave rise to the widespread species (northern temperate distribution species L. carinthiacum and L. rotatum), and the other formed the Himalayan species.A taxonomic revision of the whole genus Lomatogonium is presented. In this paper, one new section (Sect. Sarcorhizoma), one new species (L. zhongdianense S. W. Liu et T. N. Ho) and one new variety (L. forrestii var. densiflorum S. W. Liu et T. N. Ho) are described. The key to the species is given. Type studies are made for all the taxa.     

Seasonal distribution and abundance of tsetse flies (Glossina spp.) in the Faro and Deo Division of the Adamaoua Plateau in Cameroon

Ten years after the large-scale tsetse control campaigns in the important cattle rearing areas of the Faro and Deo Division of the Adamaoua Plateau in Cameroon, the seasonal distribution and abundance of tsetse flies (Glossina spp.) were determined. During a period of 12 consecutive months (January-December 2005), the tsetse population was monitored along four trap transects consisting of a total of 32 traps and two flyround transects traversing the study area, which comprised the tsetse-infested valley, a buffer zone and the supposedly tsetse-free plateau. Throughout the study period, a total of 2195 Glossina morsitans submorsitans and 23 Glossina tachinoides were captured in the traps and 1007 G. m. submorsitans (78.8% male flies) were captured along the flyround transects. All G. tachinoides and almost all G. m. submorsitans were captured in the valley. Five G. m. submorsitans were captured in traps located in the buffer zone, whereas no flies were captured in traps located on the plateau. The index of apparent abundance (IAA) of G. m. submorsitans was substantially higher in the areas close to game reserves. In the remaining part of the valley, where wildlife is scarce and cattle are present during transhumance (dry season), the IAA of tsetse was substantially lower. In this part of the valley, the abundance of tsetse seemed to be associated with the presence of cattle, with the highest IAA during transhumance when cattle are present and the lowest apparent abundance during the rainy season when cattle have moved to the plateau. It is concluded that the distribution of tsetse in a large part of the valley undergoes substantial seasonal changes depending on the presence or absence of cattle. The repercussions of those findings for the control of tsetse in the valley and the probability of reinvasion of the plateau are discussed.     

Evidence of Host Discrimination by Microctonus hyperodae Loan (Hymenoptera: Braconidae, Euphorinae), a Parasitoid of the Argentine Stem Weevil Listronotus bonariensis (Kuschel) (Coleoptera: Curculionidae)

Microctonus hyperodae is a solitary endoparasitoid of the Argentine stem weevil, Listronotus bonariensis. Early investigation into the biology of the parasitoid indicated that there was no discrimination between parasitized and unparasitized hosts. However, dissection data from two experiments were analyzed according to three mathematical models. Model I was based on the Poisson distribution and assumed random selection of hosts. The host discrimination model (model II) assumed that parasitized hosts had reduced attractiveness to searching parasitoids. A competition model (model III) made the assumption that competition between the early immature parasitoid stages led to premature mortality, which was not accounted for in the dissection results. The dissection data indicated statistically significant departure from the Poisson model. Results from the laboratory experiment indicated that host discrimination explained the results more accurately than parasitoid larval competition. Models II and III both provided adequate fits to the field data, although the departure from the observed data was greatest for the competition model. Both intra- and inter-ecotypic host recognition were evident, and there was some indication that clonal recognition took place. This is the first evidence of host discrimination by a member of the genus Microctonus. It has provided further indication of the success of M. hyperodae as a biological control agent.     

Échinodermes de l'Ordovicien supérieur (Ashgill) de Sardaigne et d'Algérie

Research on the influence of sea level variations on the benthic faunas have been carried out in the Upper Ordovician of Sardinia. Study of the depositional facies and sequence analysis of the upper part (Lower Ashgill) of the Portixeddu Formation led to the identification of the sedimentary environments. Cystoids and crinoids are associated to bryozoans and brachiopods in most levels. The numerical analysis of associations and megaguilds shows that crinoids and cystoids have a higher frequency in the proximal and median facies of the upper offshore. The columnal association characterized by Conspectocrinus celticus and the coronoid Mespilocystites tregarvanicus has been discovered in the upper part of the formation. This material and complementary samples from Upper Ordovician of Sardinia and Kabylia (Algeria) bring additional data on the systematic and show the wide distribution of this fauna outside of the Ibero-armorican domain. The distribution of this echinoderm association supports a palaeogeographical position of the Ibero-armorican domain and Sardinia within the north gondwanan margin during the Lower Palaeozoic.     

New freshwater diatom genus,Edtheriotia gen. nov. of the Stephanodiscaceae (Bacillariophyta) from south‐central China

We describe a new genus and species of the diatom family Stephanodiscaceae with light and scanning electron microscopy from Libo Small Hole, Libo County, Guizhou Province, China. Edtheriotia guizhoiana gen. & sp. nov. has striae across the valve face of varying lengths, and are composed of fine striae towards the margin and onto the mantle. Many round to stellate siliceous nodules cover the exterior of the valve. External fultoportulae opening are short tubes; the opening of the rimportula lacks a tube. Internally a hyaline rim is positioned near the margin. Marginal fultoportulae possess two arcuate opercles. Areolae have domed cribra on the valve face. Those near the margin do not open to the interior. One to three sessile to slightly‐raised rimportulae are placed on the valve face towards the margin; they do not terminate shortened striae towards the center of the valve. This suite of features distinguishes this group from other known genera in the Stephanodicaeae. Cyclotella shanxiensis is transferred to Edtheriotia, making the new combination E. shanxiensis (Xie & Qi) Kociolek et al. comb. nov. It differs from E. guizhoiana by the presence of open pores near the margin of the valve. Features of the new genus are compared and contrasted with others in the Family Stephanodiscaceae. The two species of the genus, unlike most in the family, are known only from rivers and ponds from China and Japan.     

Le Lias et le Dogger du sud du Bassin parisien (France) : rôle de la tectonique syn-sédimentaire et reconstitutions paléogéographiques à l'échelle de la biozone d'ammonite

The Lias and Dogger series in the southern Paris Basin outcrops are precisely dated following the ammonite biozonation scale. In these series, the depositional environment context is accurately identified and thus various isopic zones can bee recognized. From the outcrop study, the depositional sequence and the tectonics events are also distinguished. The poorly dated subsurface data should be not anymore interpreted as diagraphic signatures, with the traditional peak to peak correlation. They now need to be analysed according to the geodynamic evolution of each isopic zone. The Liassic transgressive series (in the Armorican continent) and the Dogger platforms numerous carbonate bodies are correlated at biostratigraphic scale, throughout the whole southern Paris Basin. Subsequently, paleogeographic maps are established and based on depositional sequences. The basin evolution image changes radically. This shows clearly that the complexes carbonate bodies layout prohibits any depositional environment interpretation and any isopach mapping at geological stage level. If not, the carbonate platforms of different age would be amalgamated, without any connection with the reality. Over the time, paleogeographic maps show syn-sedimentary short wavelength deformations, more or less well expressed according to the local sedimentary context. During the Lias and the Dogger, three steps can be distinguished in the southern Paris Basin. Hettangian to Lower Aalenian, the progressive increase in accommodation drives to total immersion of the continental areas; the maximum of accommodation during the Middle Toarcian entails uniform sedimentary conditions. However, short wavelength deformations persist. The major accidents defining the isopic zones are mainly North-South oriented. Middle Aalenian to Middle Bathonian, isopic zones are contrasted, controlled by the North-South accidents but also the North West-South East ones ; the variable accommodation allowed the development of regional sedimentary gaps and shallow limestone progradations on a marly area (so called “Sillon marneux”) ; tectonics events correspond to the Tethyan and the North Sea riftings influences. Upper Bathonian to Upper Oxfordian, the North West-South East tectonics accidents are particularly active; this tectonic phase could be associated to the thermal event, described in the Paris Basin and announcing the Malm depositional environment.     

An Ultrastructural Study on the Development of Egg- apparatus in Rice Embryo Sac

The development of the egg-apparatus (consisted of an egg cell and two synergids) of rice ( Oryza sativa L. ) was studied at the uhrastructural level. The walls of the egg cell and synergids, immediately after their formation, possessed numerous plasmodesmata. Plasmodesmata were also present on walls between the egg cell and synergids. During the enlargement phase of the egg cell and synergids, the walls at the tip region began to loosen and vesiculate. By the time the embryo sac became mature, the part of the wall of the egg cell and synergids, facing the chalaza, disappeared. Consequently, the tip regions of the egg cell and synergids were only protected by a plasma membrane. When the embryo sac reached full maturity, the upper and middle region of the wall of the synergids broke up into pieces. At that time one synergid began to degenerate. Plasmodesmata persisted at the hook region of the wall of both the egg cell and synergids. Most plastids in the egg cell contained starch grains that persisted throughout the period of the embryo sac development. Starch grains in the plastids of the synergids appeared only before the time when the two polar-nuclei moved into the region above the egg-apparatus. They then disappeared and did not appear again until the embryo sac had reached full maturity. The size and location of the vacuoles in the egg cell were different from those in the synergids. The time of formation was also different. Vacuoles in the egg cell formed late in comparison with the synergids. Vacuoles in the chalazal region of the egg cell (especially at the early stage of the embryo sac development) were much larger than those in the micropylar region. Vacuoles in the synergids tended to concentrate mainly in the chalazal region. There was a peak period of lipid formation in the two synergids. The peak appeared when the embryo sac neared maturity. At the early stage of development, the nuclei of the synergids were elliptical in shape and were situated at the central region near the micropyle. The shape of the nuclei at the late stage of development became less regular and tended to move more towards the micropylar region. Changes in the uhrastructure of the egg cell and synergids of rice appeared to be closely related to the metabolic processes controlling the embryo sac formation and development.     

A reassessment of the relations in Malaysia between ants (Crematogaster) on trees (Leptospermum and Dacrydium) and epiphytes of the genus Dischidia (Asclepiadaceae) including 'ant-plants'

Three species of epiphytic Dischidia have been investigated in terms of their relationship to ants on trees. Two species, D. parvifolia and D. astephana , are associated with ants and trees in montane areas. A clear association has been found between ants of the genus Crematogaster and the tree Leptospermum flavescens. This relationship is complex and probably both organisms benefit from the association. The ants live in tunnels in the wood of the major branches and the trunk, and the entire tree is occupied by one ant colony. Trees occupied by ants are maintained by the ants substantially clear of epiphytes other than the two species of Dischidia. The potential benefits to the tree and to the ants of this association are noted. The roots of D. astephana and D. parvifolia penetrate into the cavities of these ant nests and presumably gain nutrients from waste in the ant nests. Both Dischidia species are effectively scavenging upon the waste material from the ant-tree association. The leathery dome-shaped leaves of D. astephana are not vital to the development of the scavenging habit as D. parvifolia has lens-shaped leaves, but may offer some advantage to D. astephana by the uptake of nutrients from waste deposited by the ants under the dome-shaped leaves by interception of stem flow and by uptake of gaseous waste. Ants do not nest under these leaves. Seeds of these species of Dischidia are taken by ants into the central woody area of the ant nest where they germinate. Both Leptospermum and Dischidia can be visualized as showing adaptations to a nutient-deficient tropical montane environment. These adaptations are discussed as is the need for reassessment in this genus of the term 'ant-plant', and the need for wider recognition of the 1ant-tree' relationship between Crematogaster and Leptospermum.     

Early embryonic development of the mayfly Ephemera japonica McLachlan (Insecta: Ephemeroptera,Ephemeridae)

In the newly laid egg of the mayfly Ephemera japonica, an egg nucleus (oocyte nucleus) at metaphase of the first maturation division is in the polar plasm at the mid-ventral side of the egg, and a male pronucleus lies in the periplasm beneath a micropyle situated just opposite the polar plasm or at the mid-dorsal side of egg. The maturation divisions are typical. An extensive and circuitous migration of the male pronucleus is involved in the fertilization process: it first moves anteriad in the periplasm from beneath the micropyle to the anterior pole of the egg and then turns posteriad in the yolk along the egg's long axis to the site of syngamy, near the center of the egg. Cleavage is superficial. The successive eight cleavages, of which the first five are synchronized, result in the formation of the blastoderm, and about ten primary yolk cells remain behind in the yolk. Even in the newly formed blastoderm, the thick embryonic posterior half and the thin extraembryonic anterior half areas are distinguished: the former cells are concentrated at the posterior pole of the egg to form the germ disc, and the latter cells become more flattened, forming serosa. Time-lapse VTR observations reveal a yolk stream that is in accord with the migration of the male pronucleus in time and direction. The yolk stream is also generated in activated unfertilized eggs, and it is probable that the migration of the male pronucleus in association with the fertilization may be directed by the yolk stream. J. Morphol. 238:327–335, 1998. © 1998 Wiley-Liss, Inc.     

Pandanus subg. Martellidendron (Pandanaceae) part II: revision of sect. Martellidendron Pic. Serm. in Madagascar

This paper provides a taxonomic revision of Pandanus sect. Martellidendron , which so far comprises six species of which only one is known from both staminate and pistillate plants. Research in herbaria and in the field in Madagascar has provided the data on which a revision of the unclear taxonomy of this section can be based. Based on micromorphology, architecture and phytogeography, the number of these dioecious species is reduced. One new species (P. gallinarum Callmander) from the Biosphere reserve of Mananara-North on the east coast is described, and P. karaka Martelli is transferred to this section 30 years after it was assigned to section Dauphinensia. A key to all species of subg. Martellidendron is provided. Finally, the important role of the section for the understanding of the phylogeny of the Pandanaceae is discussed in the context of Indian Ocean biogeography.