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Enzyme
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Query: EC:2.3.1.28 (
chloramphenicol acetyltransferase
)
5,100
document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)
It has been shown that tandems of rare arginine codons
AGG
have a strong inhibitory effect on translation of mRNA in E. coli [5]. This has been explained by the rate-limiting interaction of these codons with the less abundant tRNA(
AGG
) [6]. In this study tandemly repeated
AGG
triplets were introduced into the
chloramphenicol acetyltransferase
(
CAT
) gene either upstream of the initiation ATG codon or downstream of it (both in frame and out of frame) and the expression of the modified genes was investigated. We report that the addition of
AGG
clusters resulted in a substantial inhibitory effect on
CAT
gene expression independently of their localization in mRNA. This inhibitory effect is explained by a competition of the tandem AGGAGG with the natural Shine-Dalgarno (SD) sequence (consensus AAGGAGGU) for the 3'-end of the 16S small ribosomal RNA (rRNA).
...
PMID:Effect of tandemly repeated AGG triplets on the translation of CAT-mRNA in E. coli. 137 38
Presented here is an experimental demonstration of our theoretical predictions on the role of the downstream context of unfavourable codons in a gene on its expression level. Six non clustered
AGG
codons were inserted in the
chloramphenicol acetyltransferase
(cat) gene of E. coli and the expression of this modified gene (cat4) was compared with that of a cat gene in which four clustered
AGG
codons were inserted (cat2 gene). As predicted, the rate of production of the corresponding CAT4 and CAT2 proteins is equal as long as the rate of transcription of the gene does not exceed a given limit. When this limit is exceeded, production of CAT4 continues to increase, whereas CAT2 production decreases dramatically. Various consequences and possible applications of this downstream context effect are discussed.
...
PMID:The maximum rate of gene expression is dependent on the downstream context of unfavourable codons. 251 83
The
AGG
and AGA are the least used arginine codons in E. coli but they are the most preferable ones in eukaryotes. The low expression of some eucaryotic genes (such as human alpha-1 interferon gene) which contain clusters of
AGG
codons is explained either by the limited pool of the tRNA(
AGG
) (Varenne and Lazdunski, 1986) or by the competition of these clusters with the Shine-Dalgarno (SD) sequence (Ivanov et al., 1992). The aim of the present study is to demonstrate the in vivo capacity of
AGG
tandems to bind to bacterial ribosomes. The two tandems of
AGG
codons (Arg12 Arg13 and Arg163 Arg164) of hIF alpha 1 with their surrounding nucleotides were cloned in a bacterial expression plasmid containing a strong promoter and a reporter gene (
chloramphenicol acetyltransferase
, CAT) devoid of a ribosome binding site. The results obtained showed that both
AGG
tandems initiated translation of the CAT mRNA with an efficiency equal to that of the consensus SD sequence and several fold higher than the native SD sequence of the CAT gene.
...
PMID:Domains in human interferon alpha-1 gene containing tandems of arginine codons AGG play the role of translational initiators in E. coli. 764 Oct 76
In a previous study, we have identified four types of 3' terminal codon pairs depending on their frequency of occurrence in the Escherichia coli genome: overrepresented, moderately represented, underrepresented, and missing. In this study, the influence of eight codon pairs belonging to these four groups on the efficiency of
chloramphenicol acetyltransferase
( cat) gene expression in E. coli is examined. Our results show that the missing codon pairs CCU:UAG (Pro:Stop) and CCC:UAG (Pro:Stop) had decreasing effect, whereas another missing pair CCU:
AGG
(Pro:Arg) had an opposite effect on the yield of CAT protein in comparison with the wild-type cat gene.
...
PMID:Effect of 3' terminal codon pairs with different frequency of occurrence on the expression of cat gene in Escherichia coli. 1505 75
In Escherichia coli mRNA, the arginine codons AGA/
AGG
and the isoleucine codon AUA are rarely used with frequencies of about 0.35% and 0.41%, respectively. Six genes with a different number of these codons were expressed in an E. coli in vitro coupled transcription/translation system, which contained either tRNA prepared from E. coli cells carrying a plasmid with argU and ileX genes encoding rare tRNAs (tRNA(arg)(AGA/
AGG
) and tRNA(ile)(AUA)), designated codon-plus tRNA, or normal tRNA from cells lacking the plasmid. Genes having a low number of the rare codons, such as genes encoding
chloramphenicol acetyltransferase
and anti-gp120 single-chain Fv (artificially constructed to remove rare codons), were expressed at similar levels using with both tRNA preparations. On the other hand, the use of codon-plus tRNA increased the expression levels of genes having a relatively large number of the rare codons, including genes encoding archaeal proteins, green fluorescent protein of jelly fish origin, and a single-chain Fv of mouse origin, by about 20% higher than that using normal tRNA.
...
PMID:Dosage effect of minor arginyl- and isoleucyl-tRNAs on protein synthesis in an Escherichia coli in vitro coupled transcription/translation system. 1623 46