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31.
Mitochondrial DNA was examined in natural and hatchery-reared stocks of brown trout, using different methods of restriction analysis. The methods included the development of a brown trout mt DNA hybridization probe through cloning part of the brown trout mitochondrial genome. In addition, fragments were analysed by ethidium bromide staining and end-labelling. The relative merits of each of these methods in assessing levels of genetic relatedness between the natural and hatchery-reared brown trout stocks were evaluated. In addition, the study revealed a diagnostic mtDNA restriction pattern which could be used as a genetic marker for the discrimination of these two groups of brown trout. 相似文献
32.
R.M. Kaplan J.B. Dame G.R. Reddy C.H. Courtney 《International journal for parasitology》1997,27(12):1585-1593
Accurate snail intermediate host infection prevalence data have the potential to be extremely useful in determining seasonal transmission dynamics of Fasciola hepatica. Because the microscopic techniques currently used lack the sensitivity and specificity necessary to obtain meaningful infection prevalence data, we developed a highly accurate and efficient DNA probe assay. The assay has a sensitivity of 100%, a specificity of >99%, easily detects a single miracidia and does not cross-hybridize with DNA of Fascioloides magna, Paramphistomum liorchis or Heterobilharzia americana, trematodes that share the same intermediate host and enzootic range as Fasciola hepatica. Using this assay, we determined the prevalence of F. hepatica in its snail intermediate host, Fossaria cubensis, during the second year of a 2-year study on the epizootiology of Fasciola hepatica in Florida. The overall infection prevalence of snails assayed in this study (n = 5246) was 1.5% and ranged from 0.1% to 3.1% for individual cattle ranches. Additionally, infection prevalence differed significantly for successive size groupings of snails, varying from 0% for 1-mm snails to 18.5% for 9- and 10-mm snails. The accuracy and efficiency of the DNA probe assay reported here for determining snail infection prevalence offers an inexpensive alternative to tracer animal studies for determining the epizootiology of F. hepatica. 相似文献
33.
利用Oligo1000DNA合成仪(Beckman)合成了长度为45bp的寡核苷酸单链,经纯化后用同位素γ-32P-ATP作5′末端标记后,制备成鱼类LZF-IDNA指纹探针。通过对鱼类的群体实验、亲子鉴定实验、组织细胞的稳定性实验和鱼类种类的适用范围实验后,测得:(1)LZF-IDNA指纹探针属多位点寡核苷酸探针;(2)LZF-IDNA指纹探针在鱼类种群中的鉴别机率为9.23×10-16;(3)LZF-IDNA指纹探针在鱼类亲子鉴定实验中的父系概率为0.999962;(4)LZF-IDNA指纹探针,是一种稳定的,既具有个体识别能力,又具有一定种属特异性的、适用于鱼类DNA指纹图研究的基因指纹探针。 相似文献
34.
We have developed a new method for the prediction of the lateral and the rotational positioning of transmembrane helices, based upon the present status of knowledge about the dominant interaction of the tertiary structure formation. The basic assumption about the interaction is that the interhelix binding is due to the polar interactions and that very short extramembrane loop segments restrict the relative position of the helices. Another assumption is made for the simplification of the prediction that a helix may be regarded as a continuum rod having polar interaction fields around it. The polar interaction field is calculated by a probe helix method, using a copolymer of serine and alanine as probe helices. The lateral position of helices is determined by the strength of the interhelix binding estimated from the polar interaction field together with the length of linking loop segments. The rotational positioning is determined by the polar interaction field, assuming the optimum lateral configuration. The structural change due to the binding of a prosthetic group is calculated, fixing the rotational freedom of a helix that is connected to the prosthetic group. Applying this method to bacteriorhodopsin, the optimum lateral and rotational positioning of transmembrane helices that are very similar to the experimental configuration was obtained. This method was implemented by a software system, which was developed for this work, and automatic calculation became possible for membrane proteins comprised of several transmembrane helices. © 1995 Wiley-Liss, Inc. 相似文献
35.
M. S. Ghatge V. V. Deshpande P. K. Ranjekar 《World journal of microbiology & biotechnology》1995,11(2):144-147
Restriction analysis of the genomic DNA from a high glucose/xylose-isomerase-yieldingStreptomyces sp. NCIM 2730 revealed a number of distinct bands on a background smear, indicating the occurrence of repeated DNA sequences in the genome. Optical renaturation analysis indicated that 25% of the genome comprised rapidly reannealing sequences with a copy number of 50 and a kinetic complexity of 3×103. Hybridization of theStreptomyces genomic library with theStreptomyces DNA, supported the estimate of the repetitive DNA content derived from the re-association kinetics of the DNA. Hybridization of DNA from three differentStreptomyces species with a rice repetitive DNA probe revealed the presence of homologous sequences, which is a unique finding.M.S. Ghatge was and V.V. Deshpande and P.K. Ranjekar are with the Division of Biochemical Sciences, National Chemical Laboratory, Pune -41108, India; M.S. Ghatge is now with the Department of Microbiology & Molecular Biology, The University of Kansas Medical Center, 36th and Rainbow Blvd, Kansas City, Kansas - 66103, USA. 相似文献
36.
采用1-萘胺-8-磺酸(ANS)为疏水探针,对大鼠胃粘膜表面疏水性作了研究,结果表明:以ANS(25μmol/L)与胃粘膜表面刮取物(胃粘液凝胶层)混合后的萤光强度(正常为1.23±0.19RFU/胃)可代表该粘液层的疏水性;以不同浓度ANS与胃粘液混合后的萤光强度呈饱和趋势,可用Scatchard作图法求得粘液中ANS的最大萤光强度(2.467±0.638RFU/胃)和相对亲和系数(0.032±0.016),它们可分别代表胃粘液中疏水基团的总量和单个基团的疏水性,从而可阐明胃粘膜被盐酸损伤后凝胶层粘液的ANS萤光减弱,系其疏水基团总量减少,而非单个基团的疏水性改变所致。 相似文献
37.
A highly sensitive fluorimetric assay using 3-O-methylfluorescein phosphate as substrate was used in the determination of K+-dependent phosphatase activity in preparations of rat skeletal muscle. The gastrocnemius muscle was chosen because of mixed fibre composition. Crude, detergent treated homogenate was used so as to avoid loss of activity during purification. K+-dependent phosphatase activities in the range 0.19–0.37 μmol · (g wet weight)−1 · min−1 were obtained, the value decreasing with age and K+-deficiency. Complete inhibition of the K+-dependent phosphatase was obtained with 10−3 M ouabain. Using a KSCN-extracted muscle enzyme the intimate relation between K+-dependent phosphatase activity and (Na+ + K+)-activated ATP hydrolysis could be demonstrated. A molecular activity of 620 min−1 was estimated from simultaneous determination of K+-dependent phosphatase activity and [3H]ouabain binding capacity using the partially purified enzyme preparation. The corresponding enzyme concentration in the crude homogenates was calculated and corresponded well with the number of [3H]ouabain binding sites measured in intact muscles or biopsies hereof. 相似文献
38.
Staining of living bacteria with rhodamine 123 总被引:5,自引:0,他引:5
Tohey Matsuyama 《FEMS microbiology letters》1984,21(2):153-157
Abstract It is possible to stain live bacteria with rhodamine 123 (R123). The stained fluorescent cells still keep the ability to replicate ( Staphylococcus aureus, Bordetella pertussis ) and to swim (e.g., Salmonella minnesota ). Dead cells or cells with a dissipated transmembrane potential showed markedly diminished fluorescence. Gram-negative strains were stained with different efficiency, presumably reflecting the different constitutions of the outer membrane. 相似文献
39.
Internodal and whorl (branch) cells of the green alga,Chara corallina Klein ex Willd., em. R.D.W., were studied with the extracellular vibrating probe for measuring transmembrane ion currents, and with an extracellular pH microprobe for measuring the surface pH profile. Bands of positive inward current (OH- efflux) 1–3 mm wide were separated by wider bands of outward current (HCO
3
-
influx) along the length of the cell. The measured peaks of inward current ranged from 20 to 60 A cm-2 (98 m from the cell surface) which would correspond to a surface ionic flux of 270–800 pmol cm-2 s-1. The peaks of outward current (HCO
3
-
influx) ranged from 10 to 30 A cm-2 which would correspond to a surface ionic flux of 140–400 pmol cm-2 s-1. The inward current bands matched the regions of surface alkalinity very well. The outward current (HCO
3
-
influx) was reduced at least 10-fold in low-HCO
3
-
medium, with a commensurate readjustment in the strength and pattern of inward current (OH- efflux). (Although these experiments involved a manipulation of the external pH, it is felt that the main adjustment in current patterns was in response to the reduction in exogenous HCO
3
-
). The presence of the vibrating probe perturbed the inward current region when vibrating with a 26-m amplitude, but this perturbation was eliminated when a 7-m amplitude was used. The perturbation was usually observed as a reduction in the number of inward current peaks with an increase (approximate doubling) in the amplitudes of the one or two remaining peaks. Both the inward and outward currents were light-dependent, falling off within seconds of light removal. 相似文献
40.