Gene/Protein Disease Symptom Drug Enzyme Compound
Pivot Concepts: Gene/Protein Disease Symptom Drug Enzyme Compound   Target Concepts: Gene/Protein Disease Symptom Drug Enzyme Compound

---

Query: EC:2.3.1.28 (chloramphenicol acetyltransferase)
5,100 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

Eight eukaryotic promoters have been tested for their activity in vivo in Escherichia coli. The rat beta-actin, rat amylase, rat chymotrypsin B, mouse metallothionein I, rat insulin I, human insulin, Rous sarcoma virus long terminal repeat (RSV LTR) and hepatitis B viral precore promoter activities were measured by using the bacterial chloramphenicol acetyltransferase coding sequences as the reporter function and by primer extension RNA analysis. All eight promoter-chloramphenicol acetyltransferase constructs produce chloramphenicol acetyltransferase activity with the following relative strengths: RSV LTR greater than rat beta-actin greater than rat insulin I greater than rat amylase greater than hepatitis B virus precore greater than human insulin greater than rat chymotrypsin B greater than mouse metallothionein I. A primer extension analysis indicates that transcription from the RSV LTR, rat insulin I, and rat beta-actin promoters initiates at the sites expected for eukaryotic rather than prokaryotic promoters. Thus the site of initiation is determined by the DNA sequence rather than by the RNA polymerase.
...
PMID:Eukaryotic promoters drive gene expression in Escherichia coli. 268 Nov 82

Two overlapping clones spanning 19 kilobase pairs (kb) of the 5' end of the alpha 1 (IV) collagen gene were isolated and found to contain a single exon which encoded the 5'-untranslated sequence and 84 base pairs of the signal peptide. The 5' end of this exon was determined to be the 5' end of the transcript by S1 nuclease protection and primer extension. The nucleotide sequence of 1 kb of the 5'-flanking DNA was extremely G + C-rich (greater than 70%) and contained two GC boxes and a putative cAMP regulatory sequence. The transcriptional regulation of the alpha 1 (IV) gene was studied with chimeric gene constructs utilizing 2.5 kb of the 5'-flanking sequence coupled to the gene for chloramphenicol acetyltransferase. Transfection of this construct into differentiating F9 cells resulted in low chloramphenicol acetyltransferase activity compared to beta-actin or Rous sarcoma virus long terminal repeat promoters, although these cells produce large amounts of collagen IV. Inclusion of a 2.7-kb sequence 2.3 kb downstream from the first exon in either orientation increased the transcription of the chloramphenicol acetyltransferase construct approximately 10-fold in F9 cells, but was not active in NIH 3T3 cells, which synthesize little collagen IV. These results indicate the presence of an enhancer within the first intron, which increases the expression of this gene.
...
PMID:Characterization of the promoter for the alpha 1 (IV) collagen gene. DNA sequences within the first intron enhance transcription. 284 28

Transcription of the beta-actin gene is rapidly inducible in response to serum stimulation. To determine the regions responsible for serum inducible and basal level expression, the human beta-actin promoter was subjected to mutational analysis. Two distinct elements, the CCAAT homology and the beta-actin specific conserved sequences, were found by a chloramphenicol acetyltransferase expression assay and sequence comparisons, and then analyzed for possible functions. Using a DNA bend assay, it was shown that the conserved sequences included the core of a sequence-directed bend of DNA. Gel mobility shift and DNase I protection assays revealed that the conserved sequences and the CCAAT homology were recognized by binding factors in HeLa cell extracts.
...
PMID:DNA bending and binding factors of the human beta-actin promoter. 291 21

alpha-skeletal actin message levels have been shown to be tightly regulated in chicken primary myoblast cultures. To test for gene elements required for muscle cell specific expression, DNA sequences containing the 5'-flanking regions of the chicken alpha-skeletal actin, beta-cytoplasmic actin, and the histone H2b genes were linked to the coding sequences of the chloramphenicol acetyltransferase gene and transfected into myogenic and non-myogenic cells. In contrast to beta-actin CAT hybrids, the alpha-skeletal actin CAT constructions displayed restricted CAT expression in transfected non-myogenic cells. We showed that a 411 nucleotide fragment flanking the 5' end of of the alpha-skeletal actin gene was responsible for a 9-15 fold increase in CAT enzymatic activity during myoblast fusion, versus only a transient 2 fold rise for the beta-actin and histone flanking sequences. These results indicate that DNA sequences within 411 bp of the 5' terminus of the alpha-skeletal actin gene influenced its cell type and stage specific expression.
...
PMID:Tissue restricted and stage specific transcription is maintained within 411 nucleotides flanking the 5' end of the chicken alpha-skeletal actin gene. 300 79

We present evidence that the fos oncogene encodes a transcriptional trans-activation function. trans-activation was assayed by cotransfection into NIH 3T3 mouse fibroblasts of v-fos DNA containing plasmids together with a plasmid containing a test promoter. Three v-fos DNAs were used: (i) pFBR-1, a plasmid containing the FBR proviral sequences; (ii) pFBJ-2, a plasmid harboring the FBJ proviral sequences; (iii) pMF-J, a plasmid containing the FBJ fos sequences linked to a mouse metallothionein promoter. Each of the three v-fos DNA plasmids stimulated the expression of a cotransfected chimeric gene consisting of a promoter segment of the mouse alpha 1(III) collagen gene linked to the gene for chloramphenicol transacetylase. In similar experiments the v-fos gene also stimulated the long terminal repeat promoter of Rous sarcoma virus (RSV) but neither the early promoter of simian virus 40 nor the beta-actin promoter. Evidence that the trans-activation function is specified by the v-fos coding sequences comes from the fact that a frameshift mutation in the v-fos coding sequence inhibits the trans-activation. Two mutations that map around nucleotide -100 in the RSV promoter do not respond to cotransfection with v-fos, whereas other mutations respond like the wild-type RSV promoter. These experiments suggest that the v-fos gene either encodes or induces an activator of transcription that recognizes specific sequences in promoters.
...
PMID:Transcriptional activation encoded by the v-fos gene. 301 Feb 84

Transcriptional regulation of the chicken cardiac myosin light chain 2 (MLC2-A) gene was investigated in chicken primary myoblast and fibroblast cultures transfected with vector constructs containing the bacterial marker gene for chloramphenicol acetyltransferase (CAT) under the control of the MLC2-A promoter. We here demonstrate that sequences close to the TATA box are sufficient to direct muscle specific and regulated expression of the MLC2-A mRNA. Transcription from MLC2-A promoter/CAT hybrids in myocytes starts from the authentic cap site that is also used in vivo. In primary breast muscle cells, bromodeoxyuridine (BUdR), a reversible blocking agent of cell differentiation, suppresses transcription from the MLC2-A promoter whereas nonmuscle promoters like the RSV- or the cytoplasmic beta-actin promoter are unaffected in their transcriptional capacity. Although the endogenous cardiac MLC2-A gene in chicken is exclusively active in heart, the transfected MLC 2-A promoter escapes this cell type control in primary cultures of breast muscle. These results demonstrate that although muscle specificity of the MLC2-A gene and its transcriptional up-regulation during differentiation is maintained in a rather short promoter segment, restrictive elements determining the muscle cell type specificity in vivo are either not present in our constructs or are not acting under the conditions of transient transfection.
...
PMID:The promoter of the chicken cardiac myosin light chain 2 gene shows cell-specific expression in transfected primary cultures of chicken muscle. 316 46

Approximately 1 kilobase of genomic DNA from the chicken fast myosin light-chain 1f/3f gene 5' to the transcriptional start sites for each light-chain mRNA was sufficient for differentiation-dependent, tissue-restricted expression. This was determined in primary chick myoblast cultures transfected with the chloramphenicol acetyltransferase (CAT) expression vector p8CAT containing these 5'-flanking sequences. The expression of CAT activity from both light-chain promoters was 10- to 20-fold higher in differentiated myotubes than in fibroblasts or myoblasts grown in bromodeoxyuridine. In contrast, the beta-actin and Rous sarcoma virus promoters joined to the CAT gene were expressed equally in all cell backgrounds tested. Even though the relative timing of light-chain 1f and 3f expression was altered, tissue-restricted, differentiation-dependent expression of the light-chain mRNAs was maintained with these 5' cis-acting sequence elements.
...
PMID:Approximately 1 kilobase of sequence 5' to the two myosin light-chain 1f/3f gene cap sites is sufficient for differentiation-dependent expression. 316 11

We have examined the chicken cytoskeletal actin gene for promoter activity and for the presence of cis-acting gene transcription activating sequences. Plasmids were constructed with the beta-actin promoter or other fragments of the beta-actin gene adjacent to either the truncated Herpes simplex virus (HSK)-tk promoter driving the neo gene or the enhancerless simian virus 40 (SV40) promoter driving the chloramphenicol acetyltransferase (CAT) gene. The neo plasmids were tested for frequency of transformation of mammalian cells to G418 resistance. The CAT constructions were tested for CAT expression in both transient and stable expression systems. We find that the beta-actin promoter is very strong in all of the assay systems and is as strong as any promoter we have tested. Also, we find that there are sequences in the vicinity of the beta-actin promoter which act like an enhancer sequence in activating transcription from the truncated HSV-tk promoter and the enhancerless SV40 promoter. Constructs with these actin sequences augment the transformation frequency of the neo plasmids, and stimulate the level of CAT expression from the CAT plasmids after stable chromosome insertion but not during the transient expression phase.
...
PMID:Activating elements in the promoter region of the chicken beta-actin gene. 347 Feb 37

Glucocorticoids have been shown to be useful in the treatment of certain types of chronic liver disease both by inhibiting fibrosis and by improving liver function. We have previously demonstrated in an in vivo model of hepatic fibrogenesis that dexamethasone inhibits the synthesis of types I and IV collagen. In the present study we have evaluated the level of regulation responsible for the dexamethasone-induced changes in collagen gene expression in a defined in vitro system. Primary cultures of adult rat hepatocytes treated with and without dexamethasone under classical cell culture conditions or using defined media were evaluated for synthesis and abundance of procollagen and beta-actin mRNAs. Cells treated with dexamethasone had decreased types I and IV procollagen mRNA steady state levels due in part to diminished transcription rates of the genes. On the other hand, beta-actin mRNA levels were unaffected by dexamethasone. Transient expression experiments were performed to more precisely define the mechanism whereby dexamethasone affects type I procollagen gene transcription. The recombinant plasmid, pAZ1009, containing the mouse alpha 2(I) procollagen gene promoter linked to the chloramphenicol acetyltransferase gene, was transfected into mouse fibroblast cell lines. Cells transfected with the pAZ1009 plasmid in the presence of dexamethasone had a significant decrease in chloramphenicol acetyltransferase activity when compared to cells not exposed to dexamethasone. These data suggest that dexamethasone inhibits collagen synthesis through a direct effect on the collagen gene promoter and appears also to have a post-transcriptional effect on procollagen mRNA content.
...
PMID:The effects of dexamethasone on in vitro collagen gene expression. 358 2

An efficient method of transgene modulation in fertilized eggs has been developed that uses the Cre/loxP recombination system. Twelve transgenic mouse lines carrying a chicken beta-actin promoter-loxP-chloramphenicol acetyltransferase (CAT) gene-loxP-beta-galactosidase gene construct were produced. After selection of the line showing the highest expression of the CAT gene in a variety of tissues, eggs of this line were injected in the male or female pronucleus with a Cre expression vector placed under the control of the chicken beta-actin promoter and kept in a circular form to avoid genomic integration. This resulted in a transient expression of Cre in the eggs, leading to recombination of the transgene as detected by galactosidase expression and DNA analysis. Recombination was completed before the morula stage with both types of pronuclear injections and occurred with a very high frequency; no mosaicism, no incomplete recombination, and no integration of the Cre sequence were observed in 18 mice born with this modified transgene. The beta-galactosidase gene was expressed in various tissues at levels comparable to those found for the CAT gene in the founder line. This Cre transient expression system should be useful for breeding transgenic lines in which transgene expression leads to sterility or lethality--in particular, for selecting transgenic lines with high expression of a potentially lethal transgene whose full activity is difficult to explore in a conventional transgenic system because of the risk of selecting for transgenic lines carrying only poorly expressed transgenes.
...
PMID:Site-specific recombination of a transgene in fertilized eggs by transient expression of Cre recombinase. 781 9


<< Previous 1 2 3 4 Next >>

---