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Target Concepts:
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Query: EC:3.2.1.23 (
beta-galactosidase
)
14,648
document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)
Calcium-binding protein 3 (CBP3) expression was up-regulated under the control of the actin 15 promoter and down-regulated by RNA interference in Dictyostelium discoideum. The overexpression of CBP3 accelerated cell aggregation and formed small aggregates and fruiting body. CBP3-inhibited cells showed uneven aggregation and increased slug trail lengths toward the directed light, whereas CBP3-overexpressing cells showed the opposite phenomena. Under dark condition, the enhanced slug trail length was also observed in the CBP3-inhibited cells. Yeast two-hybrid screening identified actin 8 as interacting protein with CBP3. The interaction between CBP3 and actin was confirmed by
beta-galactosidase
assay and surface plasmon resonance. CBP3 was associated with
Triton X-100
-insoluble cytoskeleton in the presence of Ca(2+) and the interaction of CBP3 with cytoskeleton was increased by the addition of Ca(2+). Using fluorescence microscopy, CBP3 was also shown to associate with the actin cytoskeleton during development. Subcellular fractionation indicated that CBP3 was enriched in cytosolic fraction. Taken together, these results suggest that CBP3 interacts with actin cytoskeleton and has a role during cell aggregation and slug migration of Dictyostelium.
...
PMID:Dictyostelium CBP3 associates with actin cytoskeleton and is related to slug migration. 1584 41
Replicative oriC plasmids were recently developed for several mollicutes, including three Mycoplasma species belonging to the mycoides cluster that are responsible for bovine and caprine diseases: Mycoplasma mycoides subsp. mycoides small-colony type, Mycoplasma mycoides subsp. mycoides large-colony type, and Mycoplasma capricolum subsp. capricolum. In this study, oriC plasmids were evaluated in M. capricolum subsp. capricolum as genetic tools for (i) expression of heterologous proteins and (ii) gene inactivation by homologous recombination. The reporter gene lacZ, encoding
beta-galactosidase
, and the gene encoding spiralin, an abundant surface lipoprotein of the related mollicute Spiroplasma citri, were successfully expressed. Functional Escherichia coli
beta-galactosidase
was detected in transformed Mycoplasma capricolum subsp. capricolum cells despite noticeable codon usage differences. The expression of spiralin in M. capricolum subsp. capricolum was assessed by colony and Western blotting. Accessibility of this protein at the cell surface and its partition into the
Triton X
-114 detergent phase suggest a correct maturation of the spiralin precursor. The expression of a heterologous lipoprotein in a mycoplasma raises potentially interesting applications, e.g., the use of these bacteria as live vaccines. Targeted inactivation of gene lppA encoding lipoprotein A was achieved in M. capricolum subsp. capricolum with plasmids harboring a replication origin derived from S. citri. Our results suggest that the selection of the infrequent events of homologous recombination could be enhanced by the use of oriC plasmids derived from related mollicute species. Mycoplasma gene inactivation opens the way to functional genomics in a group of bacteria for which a large wealth of genome data are already available and steadily growing.
...
PMID:Versatile use of oriC plasmids for functional genomics of Mycoplasma capricolum subsp. capricolum. 1593 82
Alpha-2,3-sialyltransferase (Lst) is expressed on the outer membrane of Neisseria gonorrhoeae and Neisseria meningitidis and sialylates surface lipooligosaccharide (LOS), facilitating resistance to complement-mediated killing. The enzyme is constitutively expressed from a single gene (lst) and does not undergo antigenic or phase variation. We observed that
Triton X-100
extracts of N. gonorrhoeae strain F62 contain about fivefold more sialyltransferase (Stase) activity than extracts of N. meningitidis strain MC58 [symbol: see text]3 a serogroup B acapsulate mutant. We confirmed and expanded upon this observation by showing that extracts of 16 random N. gonorrhoeae isolates contain various amounts of Stase activity, but, on average, 2.2-fold-more Stase activity than extracts of 16 N. meningitidis clinical isolates, representing several serogroups and nongroupable strains. Northern and real-time reverse transcription-PCR analysis of lst transcript levels in N. gonorrhoeae and N. meningitidis revealed that N. gonorrhoeae strains express more lst transcript than N. meningitidis strains. Although transcript levels correlate with average Stase activity observed in the two species, there was not a direct correlation between lst transcript levels and Stase activity among individual isolates of each species. Comparison of lst upstream (5'lst) regions of N. gonorrhoeae and N. meningitidis revealed striking sequence differences characteristic of the two pathogens. N. gonorrhoeae 5'lst regions possess 30-bp and 13-bp elements present as single elements or as tandem repeats that exist only as single elements in the 5'lst regions of N. meningitidis isolates. In addition, the 5'lst regions of N. meningitidis strains have 105-bp transposon-like Correia elements which are absent in N. gonorrhoeae. Chromosomal N. gonorrhoeae 5'lst::lacZ translational fusions expressed 4.75 +/- 0.09-fold (n = 4) higher
beta-galactosidase
(beta-gal) activity than N. meningitidis 5'lst::lacZ fusions in a host-independent manner, indicating differential expression is governed at least in part by sequence variations in the 5'lst regions. Reporter fusion assays and promoter-mapping analysis revealed that N. gonorrhoeae and N. meningitidis use different promoters with different strengths to transcribe lst. In N. gonorrhoeae, a strong sigma 70 promoter 80 bp upstream of the translational start site is used to transcribe lst, whereas this promoter is inactive in N. meningitidis. In N. meningitidis, a weak sigma 70 promoter at the 3' terminus of a 105-bp Correia repeat-enclosed element 99 bp upstream of the translational start site is used to transcribe lst. We conclude that differential Stase expression between N. gonorrhoeae and N. meningitidis is due at least in part to differential lst gene transcription.
...
PMID:Differential expression and transcriptional analysis of the alpha-2,3-sialyltransferase gene in pathogenic Neisseria spp. 1662
Plantaricin 423 is bactericidal to logarithmic and stationary-phase cells of Enterococcus sp. HKLHS and L. sakei DSM 20017. Detection of extracellular DNA and
beta-galactosidase
suggests that the mode of action is most probably by destabilizing of the cell membrane. Adsorption of plantaricin 423 to target cells ranged from 17% for Streptococcus caprinus ATCC 700066 to 67% for Lactobacillus plantarum LMG 13556, Lactobacillus curvatus DF38, Listeria innocua LMG 13568 and Lactobacillus sakei DSM 20017. Treatment of Enterococcus sp. HKLHS and L. sakei DSM 20017 with
Triton X-100
,
Triton X
-114 and chloroform increased the adsorption of plantaricin.
...
PMID:Parameters affecting the adsorption of plantaricin 423, a bacteriocin produced by Lactobacillus plantarum 423 isolated from sorghum beer. 1689 67
Bacteriocins ST194BZ and ST23LD, produced by Lactobacillus plantarum, inhibit Gram-positive and Gram-negative bacteria. Images obtained by atomic force microscopy showed clear signs of membrane damage of Lactobacillus sakei, accompanied by the leakage of DNA and
beta-galactosidase
. Adsorption of the bacteriocins to cells was increased when cells were treated with buffers at pH values above neutral. An increase in bacteriocin ST194BZ adsorption to cells of Enterococcus sp. and L. sakei was observed with an increase in incubation temperatures, but at different rates for the two species. Treatment of the two species with various inorganic salts and solvents gave different results regarding the adsorption of the two bacteriocins. In general, pre-treatment of the two sensitive cells with
Triton X-100
,
Triton X
-114 and chloroform increased the adsorption of the two bacteriocins. Increased adsorption of bacteriocin ST23LD to L. sakei was recorded when the cells were pre-treated with Tris and NH4-citrate. Treatment of Enterococcus sp. and L. sakei with Na-EDTA and SDS decreased the adsorption of the two bacteriocins. Variable results were recorded with inorganic salts.
...
PMID:Factors affecting the adsorption of bacteriocins ST194BZ and ST23LD to Lactobacillus sakei and Enterococcus sp. 1696 Mar 32
Bacteriocins bacJW3BZ and bacJW6BZ produced by Lactobacillus plantarum, and bacJW11BZ and bacJW15BZ produced by Lactobacillus fermentum, inhibit Gram-positive and Gram-negative bacteria. Treatment of Enterococcus sp. HKLHS and Lactobacillus sakei DSM 20017 with these bacteriocins deformed the cells and resulted in DNA and
beta-galactosidase
leakage. The bacteriocins adsorbed to sensitive and resistant strains. Optimal adsorption of bacJW3BZ and bacJW6BZ to Enterococcus sp. HKLHS was recorded at pH 10.0, whereas adsorption of bacJW11BZ and bacJW15BZ was favored at pH 4.0-8.0 and 2.0-4.0, respectively. Adsorption to L. sakei DSM 20017 was less influenced by pH. Incubation temperature had a major influence on the adsorption of bacJW6BZ and bacJW11BZ to sensitive cells, with better results recorded below 30 degrees C. Although variable results were recorded for bacJW3BZ and bacJW15BZ, optimal adsorption occurred between 37 and 60 degrees C. Variable levels of adsorption were recorded in the presence of inorganic salts and solvents, and this seems to be species-specific. Maximal adsorption (100%) was recorded for bacJW3BZ and bacJW15BZ to L. sakei DSM 20017 in the presence of most inorganic salts and solvents tested. Maximal adsorption of bacJW6BZ to Enterococcus sp. HKLHS (50%) was recorded in the presence of
Triton X
-114 and little (17%) or no adsorption in the presence of other reagents.
...
PMID:Factors affecting the adsorption of bacteriocins to Lactobacillus sakei and Enterococcus sp. 1802 82
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