人基因组百万碱基级巨型YAC的构建

人基因组百万碱基级巨型ＹＡＣ的构建戴长虹,柴建华（复旦大学遗传学研究所,上海２００４３３）关键词人基因组;酵母人工染色体（ＹＡＣ）;百万碱基;巨型ＹＡＣ人类的全部遗传信息包含在３ｘ１０＇核着酸对（ｈｐ）中,共编码５００００～１０００００个基因。人类基...     

Habitat requirements of Lycaena dispar batavus and implications for re-establishment in England

This paper reports on studies of the habitat requirements of Lycaena dispar batavus in its largest remaining site, the Weerribben National Park in the Netherlands. Observations and experiments were carried out to assess requirements for male territories, egg-laying sites and larval survival. The management of the fenland was also assessed on a landscape scale. Male territories were all situated in summer cut fen meadow. Eggs were laid on Rumex hydrolapathum in a range of habitat situations, although a preference was shown for plants on fen edges. Larval survival was highest on plants on watersides and fen edges, with no survival on plants in summer cut fen meadow. The findings are discussed in relation to whether the Broadland area in eastern England is suitable as a re-establishment site. Although many elements of the habitat mosaic are present, at the landscape scale there is insufficient open fenland and areas suitable for male territories are probably too isolated; however, a fen restoration strategy is in place which may restore suitable habitat and make re-establishment feasible in the future.     

Estimating larger carnivore numbers from track counts and measurements

Résumé Avoir conscience de l'abondance des grands carnivores est très important pour la gestion de leur conservation. Cependant, l'évaluation directe des nombres de carnivores est souvent difficile, nècessitant beaucoup d'argent et de temps, ce qui importe que l'échantillonnage indirect est une alternative fiable et valable. Les techniques de recensement à distance se sont montrèes économiques, renouvelables, et objectives. Nous avons donc estimé le nombre de grands carnivores de six espèces dans une superficie assez petite au nord de l'Afrique du Sud, en comparant trois méthodes d'échantillonnage basèes sur des décomptes des traces et mesures: (i) une approche qualitative, où les individus furent identifiés de manière subjective en étudiant les différences dans les mesures des traces; (ii) une approche quantitative qui se sert de la forte corrélation linéaire de la densité des traces à la densité des carnivores démontrée dans d'autres études; et (iii) une approche quantitative qui impliquent des analyses répétées de la variance et des post-enquêtes sur les mesures des traces afin de disposer des données statistiques pour identifier les différences entre des services des traces. Les trois méthodes fournirent des résultats comparables qui semblent raisonnables et qui peuvent être une bonne indication du vrai nombre de grands carnivores dans la zone de l'étude. Les rèsultats étayent la fiabilité des estimations indirectes de l'abondance de grands carnivores basés sur les données des traces, et sont prometteurs pour des futures recherches et pour la conservation de ces animaux.     

An acid precipitation technique: A strip assay for the large-scale DNA polymerase activity screening

A new format of a very rapid, low-cost and high-productive analysis based on the acid precipitation of radiolabeled DNA was developed. By contrast to the conventional processing of a large number of GF/C discs, the method employs one GF/C strip containing samples on individual teeth. The strip assay was validated by comparison with the glass fiber disk technique; the efficiency was demonstrated by screening E. coli superproducers and fractions obtained at the steps of Bst DNA polymerase, Large Fragment purification by the protocol we developed. The principle proposed allows simultaneous assaying many samples for the activity of different polymerases.     

Noninvasive genetic sampling reveals intrasex territoriality in wolverines

Due to its conspicuous manifestations and its capacity to shape the configuration and dynamics of wild populations, territorial behavior has long intrigued ecologists. Territoriality and other animal interactions in situ have traditionally been studied via direct observations and telemetry. Here, we explore whether noninvasive genetic sampling, which is increasingly supplementing traditional field methods in ecological research, can reveal territorial behavior in an elusive carnivore, the wolverine (Gulo gulo). Using the locations of genotyped wolverine scat samples collected annually over a period of 12 years in central Norway, we test three predictions: (1) male home ranges constructed from noninvasive genetic sampling data are larger than those of females, (2) individuals avoid areas used by other conspecifics of the same sex (intrasexual territoriality), and (3) avoidance of same‐sex territories diminishes or disappears after the territory owner's death. Each of these predictions is substantiated by our results: sex‐specific differences in home range size and intrasexual territoriality in wolverine are patently reflected in the spatial and temporal configuration of noninvasively collected genetic samples. Our study confirms that wildlife monitoring programs can utilize the spatial information in noninvasive genetic sampling data to detect and quantify home ranges and social organization.     

Complex Carbohydrate Composition of Large Dense-Cored Vesicles from Sympathetic Nerve

Highly purified noradrenergic, large, dense-cored vesicles were isolated from bovine sympathetic nerve endings by sucrose-D2O density gradient centrifugation. Their concentration of glycoprotein hexosamine and sialic acid was 6.6 and 3.9 mumol/100 mg lipid-free dry weight, respectively, values which are similar to those previously found in bovine chromaffin granules. However, whereas chromaffin granule glycoproteins are characterized by their high proportion of N-acetylgalactosamine-containing O-glycosidically-linked oligosaccharides (present in the chromogranins), such oligosaccharides accounted for only 17% of those in noradrenergic synaptic vesicle glycoproteins. Fractionation of N-3H-acetylated glycopeptides by sequential lectin affinity chromatography demonstrated that approximately two-thirds of the oligosaccharides were of the tri- and tetraantennary complex type, accompanied by 14% biantennary oligosaccharides and 3% high-mannose oligosaccharides. The vesicles had a relatively low concentration of chondroitin sulfate (less than 5% of that in chromaffin granules) but significant amounts of heparan sulfate (0.4 mumol N-acetylglucosamine/100 mg lipid-free dry weight). No hyaluronic acid was detected. The concentration of ganglioside sialic acid in the noradrenergic vesicles was approximately 1 mumol/100 mg lipid-free dry weight, which is significantly higher than that of a crude membrane mixture from which the vesicles were prepared; the ratio of N-acetyl- to N-glycolylneuraminic acid was 0.8. Several molecular species of gangliosides were detected by thin-layer chromatography, but most of these did not exactly comigrate with bovine brain gangliosides. Cholera toxin binding indicated that approximately half or less of the gangliosides belong to the gangliotetraose series.     

Preparation of antibodies that react with the large non-structural proteins of tobacco mosaic virus by usingEscherichia coli expressed fragments

Summary We have establishedEscherichia coli strains that overproduce two regions of the large non-structural proteins of tobacco mosaic virus, the 126K and 183K proteins, as fusion proteins with β-galactosidase. The two fusion proteins included respectively 514 amino acids common to both the 126K and 183K proteins, and 432 amino acids specific to the 183K protein. The synthesis of the fusion proteins inE. coli was controlled by the lipoprotein promoter andlac promoter-operator systems. After induction, the fusion proteins that were synthesized aggregated and formed inclusion bodies. Antisera raised against the purified fusion proteins reacted specifically with both the 126K and 183K proteins or with only the 183K protein in TMV-infected tobacco protoplasts. The 54K protein corresponding to the C-terminus of the 183K protein, which has been suggested to be synthesized from a third subgenomic mRNA, could not be detected by this method.     

Female size and age-specific fecundity in the small white butterfly,Pieris rapae crucivora Boisduval (Lepidoptera: Pieridae)

Individual differences in several reproductive parameters of female Pieris rapae were investigated in a controlled laboratory condition. Lifetime and age-specific fecundity showed considerable variability between individuals. Larger females began oviposition at an earlier age than smaller ones, and larger females were more fecund than smaller ones. Larger females laid a larger proportion of their eggs in the early stages of their reproductive lifetime, whilst smaller females laid the larger proportion of their eggs later in their reproductive lifetime. The significance of the variance in age-specific fecundity associated with female size is discussed with respect to the seasonal change in size and habitat utilization of this species.     

ATP-Citrate Lyase and Other Enzymes of Acetyl-CoA Metabolism in Fractions of Small and Large Synaptosomes from Rat Brain Hippocampus and Cerebellum

The activities of choline acetyltransferase and ATP-citrate lyase were significantly correlated (r = 0.995) in fractions of small and large synaptosomes isolated from rat hippocampus and cerebellum. The activities of these two enzymes did not correlate with those of pyruvate dehydrogenase, carnitine acetyltransferase, citrate synthase, acetyl-CoA synthetase, lactate dehydrogenase, or with the rate of high-affinity glutamate uptake in the synaptosomal fractions. The results provide additional evidence linking ATP-citrate lyase to the cholinergic system in the brain.     

Isolation and characterization of a virulent phage for Rhodobacter sphaeroides

A new virulent bacteriophage, termed øRsV, was isolated from a local sewage plant on the facultative phototrophic bacterium Rhodobacter sphaeroides DSM 159 as the host organism. Electron microscopic studies revealed that in general morphology phage øRsV resembles the T-even Escherichia coli phages. The host range of phage øRsV was restricted to strains of R. sphaeroides. E. coli strains B and K 12 were not infected. The phage genome was characterized on the basis of thermal denaturation profiles and restriction analyses indicating that it consists of about 160 kb of double-stranded DNA lacking cohesive ends. The G+C content was determined to be 46.8 mol%.