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31.
Development of monoclonal antibodies that identify Vibrio species commonly isolated from infections of humans, fish, and shellfish. 总被引:4,自引:3,他引:1 下载免费PDF全文
Monoclonal antibodies (MAbs) against Vibrio species that infect humans, fish, and shellfish were developed for application in rapid identifications. The pathogens included Vibrio alginolyticus, V. anguillarum, V. carchariae, V. cholerae, V. damsela, V. furnissii, V. harveyi, V. ordalii, V. parahaemolyticus, and V. vulnificus. Three types of MAbs were selected. The first important group included MAbs that reacted with only a single species. A second group comprised a number of MAbs that reacted with two, taxonomically closely related Vibrio species. For example, of 22 MAbs raised against V. alginolyticus, 6 recognized a 52-kDa flagellar H antigen common to both V. alginolyticus and V. parahaemolyticus; V. anguillarum and V. ordalii also shared antigens. A third group included three genus-specific MAbs that reacted with almost all Vibrio species but did not react with other members of the family Vibrionaceae (e.g., members of the Aeromonas, Photobacterium, and Plesiomonas genera) or a wide range of gram-negative bacteria representing many genera. This last group indicated the possible existence of an antigenic determinant common to Vibrio species. Two of these three genus-specific MAbs reacted with heat-stable antigenic determinants of Vibrio species as well as lipopolysaccharide extracted from Vibrio species. The use of the MAbs in blind tests and diagnosis of clinical isolates indicated that three different types of bacteria, viz., live, formalin-fixed, and sodium azide-killed bacteria, were detected consistently. Overall, it was found that the genus-specific MAbs were very useful for rapidly identifying vibrios in the screening of acute infections, while the species-specific MAbs and others were useful for completing the diagnosis. 相似文献
32.
Summary Better production of pro-urokinase from human cell line was observed with 5% serum containing medium than 10% or serum free medium on Cytodex II under perfusion chemostat operations, showing 0.8×10–5 (IU/daycell) of maximum productivity at 0.020 (l/h) of dilution rate in 5% serum medium, which corresponds to 800 IU/mL at this dilution rate. Conversion of pro-urokinase was reduced in the serum-containing media. 相似文献
33.
Dong-Woog Choi Jai Young Song Man-Ho Oh Jong Seob Lee Jinho Moon Se Won Suh Sang-Gu Kim 《Plant molecular biology》1996,30(5):1059-1066
A root-specific cDNA clone, PVR3, was isolated from a bean (Phaseolus vulgaris L.) root cDNA library by a differential screening procedure. The nucleotide sequence of PVR3 contains an open reading frame coding for an 11.14 kDa polypeptide of 102 amino acid residues; the first 25 amino acids correspond to the sequence characteristic of a signal peptide. Comparison of the deduced PVR3 polypeptide sequence with the polypeptide sequences of previously cloned genes indicates that PVR3 may encode a ns-LTP-like protein. Molecular modelling of the PVR3 protein predicts that it has a three-dimensional structure that is similar to the three-dimensional model determined from the maize ns-LTP. The PVR3 mRNA accumulated mainly in the roots of young seedlings. It can be detected at low levels in flowers, but it is not detected in other organs. Genomic Southern blot analysis indicates that the genomic DNA corresponding to PVR3 cDNA is encoded by a single gene or small gene family in the bean genome. 相似文献
34.
Fernando Pliego-Alfaro Mary Joy R. Monsalud Richard E. Litz Dennis J. Gray Pamela A. Moon 《Plant Cell, Tissue and Organ Culture》1996,44(1):63-70
Inhibition of mango somatic embryo growth was inducedin vitro by treatments for 4 or more weeks with abscisic acid (0–100 M ABA) with and without high osmolarity provided by mannitol (0–10%). High osmolarity and ABA significantly affected somatic embryo length, precocious germination and the production of good quality secondary somatic embryos. High osmolarity also affected root elongation. Abscisic acid was more effective in suppressing growth and development of 0.5 cm-length somatic embryos than smaller somatic embryos. Development beyond the heart stage was significantly inhibited by both ABA and mannitol treatments. The recovery of good quality somatic embryos was enhanced by high levels of ABA (100 M) with and without mannitol (0–5%). Somatic embryos that had been pulsed with ABA were partially desiccated at different relative humidities. Weight loss was affected only by relative humidity; and ABA did not enhance desiccation tolerance.Abbreviations ABA
Abscisic acid
- 2,4-D
2,4-Dichlorophenoxyacetic acid
- MM1
Mango maturation medium
- RH
Relative humidity 相似文献
35.
Yeehn Yeeh Soon Suk Kang Hye Gi Chung Mun Su Chung Myong Gi Chung 《Journal of plant research》1996,109(2):161-168
Vitex rotundifolia L.f. is a woody perennial and has sexual and asexual modes of reproduction. Allozyme study was conducted on 550 plants in
13 Korean populations. The levels of genetic variability and divergence within and among populations, respectively, are considerably
lower and higher than the mean values for woody plants with similar life history tralts. Mean percentage of polymorphic loci
(P
P), mean number of alleles per locus (A
P), and mean genetic diversity (He
P) within populations ofV. rotundifolia were: 16.7%, 1.21, and 0.047. On average, about 79% of the total variation inV. rotundifolia was common to all populations (meanG
ST=0.208). In addition, significant differences in allele frequencies among populations were found in all polymorphic loci examined
(P<0.001). On the other hand, levels of genotypic diversity within and among populations were moderate. About 44% (18/41) of
multilocus genotypes were “local genotypes” (genotypes occurring in only one population), whereas only one “widespread genotype”
(genotypes occurring in more than 75% of the populations) were detected. The mean number of multilocus genotypes per population
(G) and mean genotypic diversity index (D
G) were 8.4 and 0.74, respectively. Most common multilocus genotypes found in populations were homozygous for five polymorphic
loci. The abundance of ramets of these genets is responsible for the low levels of expected heterozygosity within populations.
The results indicate that clonal reproduction may act as an enhancer of genetic drift by reducing effective size of local
populations ofV. rotundifolia. 相似文献
36.
David McElroy Douglas A. Chamberlain Eunpyo Moon Kate J. Wilson 《Molecular breeding : new strategies in plant improvement》1995,1(1):27-37
The use of reporter genes to characterise sequence elements that act to regulate gene expression in transgenic plants has been vital to the development of foreign gene expression strategies for use in cereal transformation. ThegusA locus ofEscherichia coli, which encodes the enzyme-glucuronidase (GUS), is by far the most popular reporter gene used in plant transformation. In this paper we extend the utility of the GUS reporter gene system in cereal transformation by describing and evaluating a number of novel constructs suitable for use in direct gene transfer experiments. These plasmids are all available from the Molecular Genetic Resource Service of the Center for the Application of Molecular Biology to International Agriculture. 相似文献
37.
38.
E. Fabbri A. Capuzzo A. Gambarotta T.W. Moon 《Comparative biochemistry and physiology. Part B, Biochemistry & molecular biology》1995,112(4):643-651
Epinephrine (EPI) is thought to act by stimulating adenylyl cyclase (ACase) and cAMP production through β-adrenoceptors in the liver of more primitive vertebrates. Recent observations, however, point to an involvement of α1-adrenoceptors in EPI action, at least in some fish species. The role of the α1- and β-adrenergic transduction pathways has been investigated in rainbow trout (Oncorhynchus mykiss) hepatic tissue. Radioligand-binding assays with the β-adrenergic antagonist 3H-CGP-12177 using hepatic membranes purified on a discontinuous sucrose gradient confirmed the presence of β-adrenoceptors (Kd0.36 nM, Bmax 8.61 fmol · mg−1 protein). We provide the first demonstration of α1-adrenoceptors in these same membranes; analysis of binding data with the α1-adrenergic antagonist 3H-prazosin demonstrated a single class of binding sites with a Kdof 15.4 nM and a Bmax of 75.2 fmol · mg−1 protein. There is a straight correlation between β-adrenoceptor occupancy, ACase activation and cAMP production. On the contrary, the role of inositol 1,4,5-trisphosphate (IP3) has to be elucidated; in fact, despite the presence of specific microsomal binding sites for IP3 (Kd 6.03 nM, Bmax 90.2 fmol · mg−1 protein), its cytosolic concentration was not modulated by EPI. On the other hand, we have previously shown in American eel and bullhead hepatocytes that α1-adrenergic agonists are able to increase intracellular concentrations of IP3 and Ca2+ and to activate glycogenolysis. These data suggest a marked variation in the liver of different fish both in terms of α1-binding sites affinity and of α1-adrenoceptor/IP3/Ca2+ transduction systems. 相似文献
39.
Soo Wan Nam Byung Moon Kim Bong Hyun Chung Dae Ook Kang Jong Seog Ahn 《Biotechnology letters》1994,16(9):897-902
Summary For the secretion of human lipocortin-1 (LC-1) in yeast, a expression and secretion vector was constructed by using the promoter and signal sequence of glucoamylase gene (STA1) of Saccharomyces diastaticus. After the cDNA of human LC-1 was ligated with the secretion vector, the resulting hybrid plasmid was transformed into S. diastaticus. When the recombinant S. diastaticus was cultivated in YPD medium, LC-1 was expressed and secreted into the extracellular medium, yielding LC-1 protein at a concentration of 2.5 g/mL. 相似文献
40.
Human immunodeficiency virus type 1 Rev is required in vivo for binding of poly(A)-binding protein to Rev-dependent RNAs. 总被引:4,自引:0,他引:4 下载免费PDF全文
L H Campbell K T Borg J K Haines R T Moon D R Schoenberg S J Arrigo 《Journal of virology》1994,68(9):5433-5438
In the absence of Rev or the Rev-responsive element, the Rev-dependent human immunodeficiency virus type 1 (HIV-1) RNAs do not behave as mRNAs; rather, they exhibit nuclear defects in splicing and/or nuclear export and cytoplasmic defects in stability and translation. A translational initiation factor, eIF-5A, has recently been shown to bind specifically to the Rev activation domain. As the binding of poly(A)-binding protein 1 (PAB1) to the poly(A) tail of mRNAs is involved in both the stability and translation of cytoplasmic mRNAs, we investigated whether Rev might influence the association of PAB1 with cytoplasmic HIV-1 RNAs. Antibodies were generated against PAB1. We used these antibodies in an immunoprecipitation assay to detect specific binding of PAB1 to cytoplasmic mRNAs. We found that in the presence of Rev, PAB1 was associated with Rev-dependent and Rev-independent RNAs in the cytoplasm of transfected cells. However, in the absence of functional Rev, we found little or no PAB1 associated with Rev-dependent RNAs. These RNAs were capable of binding PAB1 in vitro. These results demonstrate that HIV-1 RNAs are defective in PAB1 association in the absence of Rev. 相似文献