Gene/Protein Disease Symptom Drug Enzyme Compound
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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)

A novel surgical technique was developed to deliver retroviral gene vectors directly to a rat liver lobe in vivo. It was observed that viral infection efficiency was enhanced by inducing hepatocyte DNA synthesis by prior partial hepatectomy. Two retroviral vectors were used to integrate specific bacterial genes: an amphotropic virus expressing the hph gene for hygromycin B phosphotransferase and an ecotropic virus expressing the lac-Z gene for beta-galactosidase. The vectors were directed to the liver by in situ selective perfusion of the posterior liver lobes with a viral suspension with inflow and outflow catheters. Male Sprague-Dawley rats were divided into three groups. Animals in the first group underwent 70% partial hepatectomy and the remnant liver lobes were allowed to regenerate for 20 hours before perfusion with the viral supernatant. Group 2 rats were perfused with viral supernatant and 2 hours later underwent 70% partial hepatectomy. Animals in the third group were perfused with the viral supernatant without partial hepatectomy. Viral transduction of hepatocytes was assessed 4 or 6 days after treatment. Hygromycin B-resistant hepatocytes were isolated from the liver remnants of rats in group 1 (21.6%) and group 2 (26.9%). No resistant hepatocytes could be detected in hepatocytes from either control rats perfused with medium alone or those from rats that did not undergo hepatectomy (group 3). In animals that received the ecotropic virus, only those that underwent hepatectomy before virus exposure (group 1) showed a small number of hepatocytes expressing beta-galactosidase in liver sections.
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PMID:In vivo regional delivery of retrovirally mediated foreign genes to rat liver cells: need for partial hepatectomy for successful foreign gene expression. 838 43

GPD regulatory sequences were used to express a phleomycin resistance gene (Sh ble) in Schizophyllum commune, resulting in high numbers of phleomycin-resistant transformants. Attempts to express heterologous genes coding for hygromycin B phosphotransferase (hph), aminoglycoside phosphotransferase (apt), beta-glucuronidase (uidA) and beta-galactosidase (lacZ) using the same regulatory sequences were not successful and no mRNA could be detected. Cloning the hph and uidA genes in an internally deleted GPD gene resulted in truncated transcripts which ended within the 5'-parts of the heterologous genes. Cloning of the same genes as transcriptional fusions downstream from the Sh ble gene also resulted in truncated transcripts ending in the 5'-parts of these heterologous genes. It is suggested that AT-rich sequences in heterologous genes might be involved in generating these truncated transcripts, thereby preventing expression in S. commune.
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PMID:Expression of heterologous genes in Schizophyllum commune is often hampered by the formation of truncated transcripts. 950 4

The echinocandin-type antimycotic mulundocandin and its derivatives are produced by the filamentous fungus Aspergillus sydowii (strain FH2551). These agents have been considered as a potential drug to treat immunocompromised patients who suffer from severe opportunistic fungal infections. In order to generate strains with a modified mulundocandin biosynthesis, we developed molecular tools for genetic engineering of A. sydowii as an alternative to conventional strain improvement procedures. For our experiments, we used strain FH2551, which was discriminated from other Aspergillus strains by determining the sequence of the two internal transcribed spacers (ITS1 and ITS2) of the rDNA locus. In addition, the electrophoretic karyotype of A. sydowii was established using pulsed-field gel electrophoresis (PFGE), leading to a calculated genomic size of about 40 Mb. For gene mapping, chromosomes were subjected to PFGE either unrestricted or after incubation with rare cutting enzymes and probed with heterologous genes. Using the bacterial hygromycin B phosphotransferase gene as a selectable marker for transformation of A. sydowii, we generated transformants with single and multiple copies of plasmid DNA. Subsequently, the heterologous lacZ and gfp genes were efficiently transferred and expressed in A. sydowii. The majority of lacZ-transformants showed more than 6 pkat beta-galactosidase activity/mg protein, while the control strains had no significant background activity. Fluorescence microscopy of gfp-transformants demonstrated that the green-fluorescent protein is present in a stable and active form in the cytoplasm of vegetative hyphae and conidiospores.
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PMID:Development of molecular tools for the mulundocandin producer Aspergillus sydowii: DNA-mediated transformation and reporter gene expression. 1195 45