Gene/Protein Disease Symptom Drug Enzyme Compound
Pivot Concepts:   Target Concepts:
Query: EC:3.2.1.23 (beta-galactosidase)
 14,648 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

An integrative cloning vector was constructed using a randomly cloned HindIII-digested chromosomal fragment from Lactobacillus acidophilus ADH inserted into an Escherichia coli vector, pBluescript II SK+. Southern hybridization studies demonstrated homology of the inserted fragment with one other L. acidophilus strain and one Bifidobacterium strain. Identification of a SauI site located near the middle of the 1.9-kb ADH chromosomal fragment made it possible to clone the Lactobacillus bulgaricus beta-galactosidase (EC 3.2.1.23) gene into this vector. The vector was unable to replicate in the homologous host, L. acidophilus ADH, following electroporation. The chromosomal fragment allowed the integration of the beta-galactosidase gene (beta gal) into the host chromosome via homologous recombination. The size of the two flanking L. acidophilus ADH chromosomal fragments, approximately 0.95 kb each, was sufficient to allow the double cross-over to take place. Southern hybridization demonstrated that only L. acidophilus and L. bulgaricus DNA had been integrated into the chromosome of the host strain. The beta-galactosidase activity of the transformant was increased approximately 200-fold when compared to the enzyme activity of the wild-type strain. The beta gal gene remained stable in the transformant strain after 30 transfers in growth media without selection pressure. This first-generation integrative cloning vector is constructed solely of DNA from organisms consumed by humans and could be considered a food-grade vector system.
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PMID:Construction of an integrative food-grade cloning vector for Lactobacillus acidophilus. 873 72

An efficient method is described for the generation of site-specific chromosomal integrations in Lactobacillus acidophilus and Lactobacillus gasseri. The strategy is an adaptation of the lactococcal pORI system (K. Leenhouts, G. Venema, and J. Kok, Methods Cell Sci. 20:35-50, 1998) and relies on the simultaneous use of two plasmids. The functionality of the integration strategy was demonstated by the insertional inactivation of the Lactobacillus acidophilus NCFM lacL gene encoding beta-galactosidase and of the Lactobacillus gasseri ADH gusA gene encoding beta-glucuronidase.
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PMID:Efficient system for directed integration into the Lactobacillus acidophilus and Lactobacillus gasseri chromosomes via homologous recombination. 1152 48

All- trans-retinoic acid (RA) contributes to the establishment of the anterior-posterior (AP) axis in chordates. In vertebrates, all- trans-retinol is oxidized to RA by two oxidative steps. However, the controversy about the enzymes responsible for retinol oxidation (ADH vs RDH) and the fact that some candidates are absent in cephalochordates questioned retinol oxidation in this lineage. Retinoid quantitation has revealed that Branchiostoma floridae adults contain both retinol and retinoic acid as well as retinal, the intermediate in the metabolic pathway. Furthermore, our data show that the developmental effects of retinol treatment are comparable to those reported for RA. SEM analysis revealed mouth and gill slit aberrations due to a posteriorization effect, also visualized by changes in the beta-galactosidase pattern. Overall, these findings support the idea that amphioxus metabolizes endogenous retinol to retinoic acid and suggest a common oxidative pathway for RA in the chordate phylum.
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PMID:Retinoic acid synthesis in the prevertebrate amphioxus involves retinol oxidation. 1220 95

Whole-mount detection methods are quick, inexpensive and offer the possibility of studying the temporal and spatial patterns of gene expression in a morphological context. These methods have been used widely to detect messenger RNAs and to measure enzymatic activity of reporter genes, such as beta-galactosidase or beta-glucuronidase. Taking advantage of the fact that NADH generated during the oxidation of formaldehyde by class III alcohol dehydrogenase can reduce the compound nitroblue tetrazolium to form a blue precipitate, we have developed a new method to detect class III alcohol dehydrogenase activity in situ in whole Arabidopsis plants. This reaction has been used earlier for in situ electrophoresis detection and for histochemical analysis in animal tissue sections. With a few modifications, it can be used in whole Arabidopsis plants or excised plant tissues to allow a rapid analysis of class III ADH activity during development or in response to elicitors. The method might be extended to other dehydrogenases by using specific substrates.
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PMID:Histochemical assay to detect class III ADH activity in situ in Arabidopsis seedlings. 1551 10