鸟分枝杆菌PhoR基因的克隆及诱导表达

目的:克隆鸟分枝杆菌的PhoR基因,构建高表达重组质粒pGEX-PhoR,确定目的蛋白表达形式,为后续对鸟分枝杆菌PhoR的功能研究打下基础。方法:以一株鸟分枝杆菌临床分离株作为DNA模板,PCR扩增PhoR基因,pGEX-4T-3为载体构建表达质粒,转化到E.coliBL21(DE3)中诱导表达。通过聚丙烯酰胺凝胶电泳(SDS-PAGE)分析蛋白表达形式及表达量。结果:扩增出鸟分枝杆菌PhoR基因,构建了重组表达质粒pGEX-PhoR,在BL21(DE3)中以可溶性方式高效表达,经诱导产生与预期值相符的一个约81KDa的融合蛋白。结论:成功构建了pGEX-PhoR重组表达质粒,在大肠杆菌中诱导表达,为后续深入研究鸟分枝杆菌PhoR的生物活性和免疫活性奠定了基础。     

假单胞菌H78中PhoR/B系统对磷元素吸收及其藤黄绿菌素合成的调控

【目的】研究根际荧光假单胞菌(Pseudomonas protegens)H78中双组分系统PhoR/B对Pst磷转运系统以及Plt生物合成的调控作用。【方法】通过同源重组的方法敲除pho R和pho B基因;使用lac Z报告基因融合质粒研究PhoR/B系统对Pst磷转运系统表达的调控;在不同磷浓度下测定H78野生型及H78pho BR突变株的生长,并在KMB培养基中测定其Plt产量。【结果】H78野生型中pst S′-′lac Z融合质粒表达的Lac Z酶活是H78pho BR突变株的15倍;在磷饥饿条件下H78的生长是H78pho BR菌株的3倍;在KMB培养基中,H78的Plt产量为H78phoBR菌株的2倍。【结论】PhoR/B系统正调控Pst转运系统的表达;在磷饥饿条件下,PhoR/B促进磷元素的吸收和利用;PhoR/B同时对Plt生物合成存在一定程度的正调控作用。     

Did the evolution of the phytoplankton fuel the diversification of the marine biosphere?

We discuss the possible links between the fossil record of marine biodiversity, nutrient availability and primary productivity. The parallelism of the fossil records of marine phytoplankton and faunal biodiversity implicates the quantity (primary productivity) and quality (stoichiometry) of phytoplankton as being critical to the diversification of the marine biosphere through the Phanerozoic. The relatively subdued marine biodiversity of the Palaeozoic corresponds to a time of relatively low macronutrient availability and poor food quality of the phytoplankton as opposed to the diversification of the Modern Fauna through the Mesozoic–Cenozoic. Increasing nutrient runoff to the oceans through the Phanerozoic resulted from orogeny, the emplacement of Large Igneous Provinces (LIPs), the evolution of deep-rooting forests and the appearance of more easily decomposable terrestrial organic matter that enhanced weathering. Positive feedback by bioturbation of an expanding benthos played a critical role in evolving biogeochemical cycles by linking the oxidation of dead organic matter and the recycling of nutrients back to the water column where they could be re-utilized. We assess our conclusions against a recently published biogeochemical model for geological time-scales. Major peaks of marine diversity often occur near rising or peak fluxes of silica, phosphorus and dissolved reactive oceanic phosphorus; either major or minor ⁸⁷Sr/⁸⁶Sr peaks; and frequently in the vicinity of major (Circum-Atlantic Magmatic Province) and minor volcanic events, some of which are associated with Oceanic Anoxic Events. These processes appear to be scale-dependent in that they lie on a continuum between biodiversification on macroevolutionary scales of geological time and mass extinction.     

Is the Octomacridae the sister family of the Diplozodae?

Diplozoidae and Octomacridae are usually considered as sister families. Essentially this is because they are the only polyopisthocotyleans parasitising primary freshwater teleosts. Because of the lack of phylogenetically informative morphological characters to explore the pattern of colonisation of the primary continental freshwater teleosts and in order to understand the appearance of the "natural parabiosis" of Diplozoidae, a molecular phylogeny was inferred by comparing newly obtained partial 28S and 18S rDNA gene sequences of Eudiplozoon nipponicum and Diplozoon homoion with other already available sequences. The phylogenetic analysis seems to show that Diplozoidae and Octomacridae are not sister groups. Thus, the colonisation of primary freshwater teleosts by these two families could be independent.     

Imperfect Batesian mimicry—the effects of the frequency and the distastefulness of the model

Batesian mimicry is the resemblance between unpalatable models and palatable mimics. The widely accepted idea is that the frequency and the unprofitability of the model are crucial for the introduction of a Batesian mimic into the prey population. However, experimental evidence is limited and furthermore, previous studies have considered mainly perfect mimicry (automimicry). We investigated imperfect Batesian mimicry by varying the frequency of an aposematic model at two levels of distastefulness. The predator encountered prey in a random order, one prey item at a time. The prey were thus presented realistically in a sequential way. Great tits (Parus major) were used as predators. This experiment, with a novel signal, supports the idea that Batesian mimics gain most when the models outnumber them. The mortalities of the mimics as well as the models were significantly dependent on the frequency of the model. Both prey types survived better the fewer mimics there were confusing the predator. There were also indications that the degree of distastefulness of the model had an effect on the survival of the Batesian mimic: the models survived significantly better the more distasteful they were. The experiment supports the most classical predictions in the theories of the origin and maintenance of Batesian mimicry.     

Phytochemicals: the good, the bad and the ugly?

Phytochemicals are constitutive metabolites that enable plants to overcome temporary or continuous threats integral to their environment, while also controlling essential functions of growth and reproduction. All of these roles are generally advantageous to the producing organisms but the inherent biological activity of such constituents often causes dramatic adverse consequences in other organisms that may be exposed to them. Nevertheless, such effects may be the essential indicator of desirable properties, such as therapeutic potential, especially when the mechanism of bioactivity can be delineated. Careful observation of cause and effect, followed by a coordinated approach to identify the responsible entities, has proved extremely fruitful in discovering roles for phytochemical constituents. The process is illustrated by selected examples of plants poisonous to animals and include the steroidal alkaloid toxin of Veratrum californicum (Western false hellebore), piperidine alkaloids of Lupinus species (lupines), and polyhydroxy indolizidine, pyrrolizidine and nortropane alkaloids of Astragalus and Oxytropis species (locoweeds), Castanospermum australe (Moreton Bay chestnut) and Ipomoea species (morning glories).     

Effect of cortisone on the developmental pattern of the neutral and the acid β-galactosidases of the small intestine of the rat

1. The developmental pattern and effect of cortisone on acid beta-galactosidase and neutral beta-galactosidase were studied in postnatal rats by a recently proposed method for their independent determination. 2. After birth the acid beta-galactosidase activity increases in the ileum, whereas it decreases slightly in the jejunum. On day 16 after birth the activity in the ileum decreases and in 20-day-old rats activity in both parts of the intestine decreases to adult values. In suckling animals the activity in the ileum exceeds the jejunal activity severalfold and in adult animals the activity in the jejunum is slightly higher than that in the ileum. 3. Neutral beta-galactosidase activity is high after birth and decreases in both jejunum and ileum after day 20 after birth. In 12-20-day-old rats activity in both parts is essentially the same, but in adult animals jejunal activity exceeds ileal activity four-to five-fold. 4. Cortisone (0.5, 2.0 or 5.0mg/100g body wt. daily for 4 days) does not influence the activity of either enzyme in 60-day-old rats. Acid beta-galactosidase activity is decreased after cortisone treatment in 8-, 12-, 16-and 18-day-old rats, with sensitivity to cortisone increasing with the approach of weaning. No effect of cortisone on acid beta-galactosidase is seen in 8-day-old rats. Neutral beta-galactosidase activity is increased in the ileum of 8-, 12-, 16- and 18-day old rats, but only in the jejunum of 8-and 12-day-old rats.     

Is Mislocalization during Saccades Related to the Position of the Saccade Target within the Image or to the Gaze Position at the End of the Saccade?

A stimulus that is flashed around the time of a saccade tends to be mislocalized in the direction of the saccade target. Our question is whether the mislocalization is related to the position of the saccade target within the image or to the gaze position at the end of the saccade. We separated the two with a visual illusion that influences the perceived distance to the target of the saccade and thus saccade endpoint without affecting the perceived position of the saccade target within the image. We asked participants to make horizontal saccades from the left to the right end of the shaft of a Müller-Lyer figure. Around the time of the saccade, we flashed a bar at one of five possible positions and asked participants to indicate its location by touching the screen. As expected, participants made shorter saccades along the fins-in (<–>) configuration than along the fins-out (>–<) configuration of the figure. The illusion also influenced the mislocalization pattern during saccades, with flashes presented with the fins-out configuration being perceived beyond flashes presented with the fins-in configuration. The difference between the patterns of mislocalization for bars flashed during the saccade for the two configurations corresponded quantitatively with a prediction based on compression towards the saccade endpoint considering the magnitude of the effect of the illusion on saccade amplitude. We conclude that mislocalization is related to the eye position at the end of the saccade, rather than to the position of the saccade target within the image.     

Does the weight of the brain depend on the body height?]

Brains of 1664 subjects (895 males and 769 females) aged from 20 to 89 years have been studied. The whole material being investigated was divided, within sex groups, into body-height classes and age classes. The class interval within the age classes was 10 years, that in height classes 5 cm. Mean arithmetics, standard deviations, standard error as well as coefficients of variation and correlation for respective classes have been calculated. It has been ascertained that the brain weight depends on the body height. In tall subjects no brains of extremely low absolute weight are encountered and, adversely, high brain weight is seldom met in short individuals. The body height also exerts certain influence upon the relative weight of the brain. More favourable proportion between the brain weight and the body length has been revealed in short subjects. Tall individuals are characterized by a low relative weight of the brain. It should be supposed that the spinal cord weight is higher in the latter subjects. The differences between the mean absolute weight of women's brains and that in men of the same age class are conditioned by the difference in the body length. A constant magnitude of difference in the mean brain weight in subjects of the same body height claims 100 g. The paper provides 2 enclosed tables representing obtained results for arithmetic mean of the absolute brain weight both in the age classes and body height classes. The differences between the mean weights of brains in women as well as in men are not significant. The coefficient of correlation between the brain weight and the body height is for men r male1 = 0.2008 for women r female1 = 0.2630, wherease the coefficient of regression for the brain weight is r male2 = 3.67 and r female2 = 3.906 respectively.     

Does DNA alone determine the development of the organism? (the information aspect of the problem)

Two closely related controversial problems are discussed: whether the developmental processes can be reduced to the synthesis of polypeptides encoded in DNA, and whether the information in DNA is equivalent to that in the adult organism. Critically considered are the ideas that DNA is only responsible for the protein synthesis, whereas morphogenesis proceeds independently and according to epigenetic regularities of its own. It is stated that development is the realization of genetic information in which more elementary (molecular) processes unambiguously determine a more complex cellular level which in its turn determines morphogenesis of tissues and organs. Various mechanisms of the appearance of new information in the course of development are considered. The statement is made that new information concerns only some individual characters of the organism, whereas most of information that determines the process of development and the structure of the adult organism is created in the course of evolution, is stored in DNA and inherited.