二核苷酸重复多态性的非同位素检测及其在基因诊断中的应用

本文报道了一种检测二核苷酸重复多态性的简便的非同位素法,利用重复序列两侧的特异引物进行ＰＣＲ扩增,产生的等位片段在薄层变性聚丙烯酰胺凝胶电泳上分离,再用灵敏的银染法显色。该方法不需要标记ＰＣＲ产物,简便、快速,分辨率可达１ｂｐ,并可用多对引物同时进行多重ＰＣＲ分析。用此方法对ＤＭＤ家系成员ｄｙｓｔｒｏｐｈｉｎ基因的５＇－脑型外显子止游区和３＇－非翻译区的两个（ＣＡ）。位点进行了扩增片段长度多态性分     

Immunoreactivity between a monoclonal lupus autoantibody and the arginine/aspartic acid repeats within the U1-snRNP 70K autoantigen is conformationally restricted

Immunoreactivity of the arginine/aspartic acid (RD) repeats of the 70K protein of U1 small nuclear ribonucleoprotein (snRNP) was determined to be conformationally dependent. The monoclonal autoantibody 2.73, isolated from a lupus-prone MRL/n mouse model, is reactive with the RD repeat regions of U1 snRNP 70K protein. Immunochemical analysis of the antigenic determinants with use of chemically synthesized peptides characterized the 2.73 epitope as the RD repeat [Pelsue, S.,et al. (1993)Autoimmunity,15, 231–236] Analysis by circular dichroism (CD) and nuclear magnetic resonance spectroscopy indicates conformational preferences in the immunoreactive peptides. Computer analyses of CD spectra obtained on the RD-containing peptides predict-turns and-sheet to be the preferred conformations of the RD repeats. This structure was also predicted by the Chou-Fasman algorithm. The RD repeat is believed to be a conserved structural motif; however, the biological function is still unclear. Immunological and biochemical analysis of autoimmune antibodies and their respective antigenic determinants has helped to characterize the possible mechanisms that lead to autoimmune diseases. This is the first report of a conformationally dependent, linear epitope of an autoantibody.     

A panel of bovine, ovine and caprine polymorphic microsatellites

We report a set of six new bovine microsatellite polymorphisms based on (CA)_n repeats. They are highly polymorphic and thus represent valuable markers for genome mapping. Four of the six are polymorphic in sheep and two are polymorphic in goats. One, which is polymorphic in cattle and sheep and apparently monomorphic in goats, is X-chromosome specific and has potential value in, for example, sex determination and detection of chimaerism.     

Effects of sexual differences of age at maturity and survival on population sex ratio

Summary Five demographical factors influencing the sex ratio of a population are classically considered. The influence of two of them is dependent on the longevity of individuals in the population. The effect of differential age at maturity between males and females is higher for animals with low annual survival, whereas the effect of differential annual survival between males and females is higher for animals with high annual survival. Such a conclusion applied to turtles, which are long life-span animals, allows us to retain differential survival between sexes as a major factor influencing the population sex ratio.     

Woody plant encroachment in granite barrens on the Frontenac Arch,eastern Ontario,Canada

Aims

Woody plant encroachment is a widespread phenomenon affecting treeless or sparsely treed habitats. We aimed to determine the extent and timing of tree and shrub encroachment into rock barrens of eastern Ontario over the last century, and to assess implications for their ongoing management.

Location

Queen's University Biological Station in the Frontenac Arch ecoregion.

Methods

We quantified the extent of change in woody vegetation in 290 rock barrens using aerial photography from 1925, 1965, and 2008. Composition and structure of woody plant communities in 10 barrens was subsequently quantified in the field using plot-based sampling. Cores or cross-sections were obtained from individuals >1.5 m height and dendrochronological techniques were used to determine their age and identify temporal patterns of any woody encroachment.

Results

Aerial photography indicated that the mean proportion of woody plant cover in barrens increased 22.5% from 1925 to 2008. Dendroecological analysis supported this. Few trees were present prior to 1900 and most established since 1960. Fraxinus americana, Juniperus virginiana, and Juniperus communis were the most common woody species colonizing the barrens. Remnants of large Pinus strobus stumps with extensive charring were found in 90% of the sampled barrens at a mean density of 22.6 stumps ha⁻¹.

Conclusions

Rock barrens on the Frontenac Arch have changed substantially over the past century; gradually being colonized by trees and shrubs and losing their distinctly open character. Active management — including prescribed fire and mechanical thinning — may be necessary if there is a desire to maintain these barrens and the rare species they support as components of the region's biodiversity. However, identification of a reference state for restoration is complicated by the fact that the structure and composition of these habitats were undoubtedly altered by European land clearance in the 19th century, and that some of these areas likely existed as pine woodlands before that.     

The Gene Encoding the Heat-Stable Enterotoxin of Vibrio cholerae Is Flanked by 123-Base Pair Direct Repeats

The heat-stable enterotoxin (O1-ST) gene (sto) was cloned from chromosome of the strain GP156 of Vibrio cholerae O1 (Inaba, El Tor) in Escherichia coli K-12, and its nucleotide seqence was determined. The nucleotide sequence of sto was very similar to that of NAG-ST gene (stn) of V. cholerae non-O1. Both sto and stn were flanked by 123-base pair direct repeats which had at least 93% homology to one another and included some inverted repeats. All the strains of V. cholerae, V. mimicus, V. metschnikovii, V. hollisae and Yersinia enterocolitica examined by colony hybridization had the direct repeat sequence regardless of ST-gene possession.     

Ribosomal DNA variation and its phylogenetic implication in the genusBrachypodium (Poaceae)

The structure of ribosomal DNA ofBrachypodium and several other grass species was investigated using a heterologous rDNA probe from wheat. Several different rDNA families were present among perennial and annual species within the genus. In contrast to the annual species the perennial species exhibited a very low degree of repeat length variation. An extra Eco RI site and a Hin dIII site were observed in the IGS, which distinguishedBrachypodium from other grass genera. The restriction fragment length polymorphism and length variation of the repeat units have taxonomic value withinBrachypodium and are correlated with the classification ofBrachypodium derived from other data.     

Selective incorporation of 5-hydroxytryptophan blocks long range electron transfer in oxalate decarboxylase

Oxalate decarboxylase from Bacillus subtilis is a binuclear Mn-dependent acid stress response enzyme that converts the mono-anion of oxalic acid into formate and carbon dioxide in a redox neutral unimolecular disproportionation reaction. A π-stacked tryptophan dimer, W96 and W274, at the interface between two monomer subunits facilitates long-range electron transfer between the two Mn ions and plays an important role in the catalytic mechanism. Substitution of W96 with the unnatural amino acid 5-hydroxytryptophan leads to a persistent EPR signal which can be traced back to the neutral radical of 5-hydroxytryptophan with its hydroxyl proton removed. 5-Hydroxytryptophan acts as a hole sink preventing the formation of Mn(III) at the N-terminal active site and strongly suppresses enzymatic activity. The lower boundary of the standard reduction potential for the active site Mn(II)/Mn(III) couple can therefore be estimated as 740 mV against the normal hydrogen electrode at pH 4, the pH of maximum catalytic efficiency. Our results support the catalytic importance of long-range electron transfer in oxalate decarboxylase while at the same time highlighting the utility of unnatural amino acid incorporation and specifically the use of 5-hydroxytryptophan as an energetic sink for hole hopping to probe electron transfer in redox proteins.