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Query: EC:3.2.1.31 (beta-glucuronidase)
 7,680 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

Polycistronic mRNAs containing an upstream beta-glucuronidase (GUS) and a downstream chloramphenicol acetyltransferase (CAT) reporter open reading frame (ORF) were expressed in transfected plant protoplasts. CAT expression could be strongly induced by coexpression of the cauliflower mosaic virus encoded translation transactivator. Transactivation was abolished when an upstream ORF overlapped the CAT ORF for a long distance. No specific sequence elements were required for transactivation but the presence of a short ORF upstream of the GUS ORF strongly enhanced the process. The inhibitory effect of additional presumed stem structures inserted into various regions of the reporter mRNAs indicates that both ORFs are translated by ribosomes that associate with the RNA at the 5' end and reach the ORFs by a linear migration mechanism.
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PMID:Translation of a polycistronic mRNA in the presence of the cauliflower mosaic virus transactivator protein. 193 8

The tetracycline-controlled transactivator (tTA)-mediated gene activation system was examined in virus infected cells to determine its role in the control of gene expression. In the presence of tTA, the gene expression from the tetO-modified minimal promoter was efficiently activated in the uninfected cells, whereas essentially no activation was observed from the only minimal promoter without the seven direct repeats of 42 bp tetO sequences. However, essentially no activation was observed when only the minimal promoter was used, without the seven direct repetitions of the 42 bp tetO sequences. On the other hand, in the infected cells, a substantial background of beta-glucuronidase expression was detected in the absence of tTA, even though tTA stimulated the gene expression by approximately 7-fold. This background expression indicates that the sequences within or nearby tetO are involved in the background stimulation of the gene expression by HCMV and HSV-1. These results suggest that the application of the tTA-mediated gene activation system may not be extremely useful for studying the biological roles of HCMV and HSV genes in the viral replicative cycles, because of the basal activity of the gene expression.
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PMID:Evaluation of tTA-mediated gene activation system on human cytomegalovirus and herpes simplex virus type-1 infections. 1089 59

The oomycete genus Phytophthora includes many important plant pathogens for which extensive genome data exist, but lacking is an inducible expression system to study contributions of their genes to growth and pathogenicity. Here the adaptation of the reverse tetracycline transactivator (rtTA) system to P. infestans is described. Vectors were developed containing rtTA expressed from an oomycete promoter, and beta-glucuronidase (GUS) controlled by TetR binding sites fused to a minimal oomycete promoter. Transformants were obtained in which GUS was expressed in a dose-dependent manner by the rtTA inducer doxycycline, indicating that the gene switch functions in P. infestans. However, toxicity of rtTA hindered the isolation of transformants if expressed on the same plasmid as the nptII selection marker. Better results were obtained by cotransforming those genes on separate plasmids, with 92% of transformants acquiring both DNAs although only 4% expressed rtTA at detectable levels. Low levels of reporter activity were measured in such transformants, suggesting that rtTA activated transcription weakly. Also, significant variation in the sensitivity of isolates to doxycycline and tetracycline was observed. These results are useful both in terms of developing tools for functional genomics and understanding the fate of DNA during Phytophthora transformation.
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PMID:Performance of a tetracycline-responsive transactivator system for regulating transgenes in the oomycete Phytophthora infestans. 1737 92