SEARCH · PubMed Health

Results for “Operon”

Explore indexed PubMed citations for clinical trials, systematic reviews and public health research. Read source abstracts and follow each citation to its original PubMed record.

Quote a phrase for an exact phrase match. Source license links do not imply unrestricted reuse.

At least 1,045 records · Page 58Linked to original sources

PubMed2002

High-level expression of foreign genes via multiple joined operons and a new concept regarding the restricted constant of total amount of plasmid DNA per Escherichia coli cell.

OBJECTIVE: To examine the feasibility of linking operons in tandem to enhance expression of heterologous genes in Escherichia coli (E. coli) and clarify the potential control mechanism of the total plasmid DNA amount in each host cell. METHODS: Two series of expression plasmids, CW11 and CW12, containing 1 to 4 and 1 to 3 heterologous gene operon(s) respectively, were constructed. The molecular size of the CW11 series varied from 5.47 kb to 12.26 kb in 2.25 kb increments. The CW12 series varied from 5.40 kb to 9.72 kb in 2.16 kb increments. The expression level of desired protein was assayed by SDS-PAGE and laser density scanning. Plasmid copy number was determined by incorporation with (3)H-thymidine ((3)H-TdR). RESULTS: No influence of the tandem-joined operons on host growth and plasmid stability was observed. Upon induction, the desired protein accumulations in the CW11 series were 44.9% +/- 3.9%, 51.3% +/- 4.1%, 54.8% +/- 3.3% and 58.2% +/- 3.4% of total cell protein. In the CW12 series, the yields were 32.2% +/- 5.0%, 42.8% +/- 4.1% and 46.9% +/- 4.0% of total cell protein. As size increased, the plasmid copy number decreased, but target gene dosage increased significantly (P < 0.01). Further calculation showed that the total amount of plasmid DNA per cell was not significantly different in each series (P > 0.05) and restricted to some extent. CONCLUSIONS: Increasing the target gene dosage by tandem linking of operons may enhance the expression level of a desired protein. Although the size (kb) and the copy number of each plasmid are negatively interrelated, for certain plasmids in each series, their total DNA amount per cell seems to be a restricted constant for specific E. coli strains under identical incubation condition.

DNA↗

PubMed2000

[High level expression of foreign gene via multiple joined operons and a new concept on the restricted constant of total amount of plasmid DNA per cell of Escherichia coli].

OBJECTIVE: To examine the feasibility of linking operons in tandem to enhance expression of heterologous genes in Escherichia coli and to clarify the potential control mechanism of the total plasmid DNA amount in each host cell. METHODS: Two series of expression plasmids, CW11 and CW12, containing 1 to 4 and 1 to 3 heterologous gene operon(s), respectively, were constructed. The molecular size of the CW11 series varied from 5.47 kb to 12.26 kb in 2.25 kb increments. The CW12 series varied from 5.40 kb to 9.72 kb in 2.16 kb increments. Protein expression was analyzed via SDS-PAGE and laser scanning. Plasmid copy number was determined by incorporation with 3H-TdR([3H]-thymidine). RESULTS: No influence of the tandem-joined operons on host growth and plasmid stability was observed. Upon induction, the desired protein accumulated in the CW11 series were 46.0%, 54.8%, 56.1%, and 60.1% of total cell protein. In the CW12 series, the yields were 33.5%, 44.0%, and 47.1% of total cell protein. With increasing size, however, plasmid copy number decreased. Further calculation showed that the total amount of plasmid DNA per cell is restricted to some extent. There seems exists a mechanism to control the maximum level of total plasmid DNA amount in each host cell. CONCLUSIONS: Increasing the target gene dosage by tandem linking of operons enhanced the expression level of a desired protein. Although the size (kb) and the copy number of each plasmid are negatively interrelated, it seems that as for certain plasmids, their total DNA amount per cell is a restricted constant for a specific E. coli strain under identical incubation condition.

Cloning, Molecular↗

PubMed1992

The alpha-operon equivalent genome region in the extreme halophilic archaebacterium Haloarcula (Halobacterium) marismortui.

The genome region of the extreme halophilic archaebacterium Haloarcula marismortui equivalent to the alpha-operon of Escherichia coli has been characterized. In H. marismortui, the alpha-operon was found to be located immediately upstream from the S9 gene cluster. The gene order in the halobacterial alpha-operon, given according to the gene products, is tRNA(Ser), HmaS13, HmaS4, HmaS11, and HmaRp alpha. Compared to the corresponding operon from E. coli, the halobacterial gene organization differs in (i) the presence of a gene for tRNA(Ser) (GCU), (ii) the reversed order of the genes for the ribosomal proteins HmaS11 and HmaS4, and (iii) the absence of the gene coding for the ribosomal protein L17. The primary structure of HmaRp alpha shows high similarity to a subunit of eukaryotic RNA polymerase II (YeaRpB3, HsaRpB33), whereas the similarity to the eubacterial alpha-subunit of RNA polymerase is only weak.

Amino Acid Sequence↗

PubMed2004

[Involvement of sigma S and sigma 70 subunits of RNA polymerase and the CRP protein in the regulation of microcin C51 operon expression].

Expression of the microcin C51 operon in Escherichia coli cells is activated during cell entry into the stationary growth phase and depends on the sigmaS subunit of RNA polymerase (RpoS). The null rpoS mutations retained the residual expression level of the transcriptional P(mcc)-lac fusion, which indicates that other sigma subunit can participate in the regulation of transcription of the microcin C51 operon. Data presented in this work show that the overproduction of sigma70 in rpoS- cells diminished the level of P(mcc)-lac expression, as in wild-type cells, which seems to be the consequence of competition between sigma factors for a limited number of core RNA polymerase molecules. In the presence of the rpoD800 mutation that renders sigma70 temperature-sensitive, expression of P(mcc)-lac was not induced in the phase of delayed culture growth at nonpermissive temperature, which indicates that sigma70 is indispensable for microcin operon expression. Point substitutions in the -10 P(mcc) region, leading to the formation of 5'-TGaTATAAT-3' site, enhanced promoter activity but did not affect the relationship between P(mcc)-lac transcription and growth phase, sigmaS, and the activator protein CRP. The activator protein CRP was shown to bind a DNA fragment containing the TGTGA(AATGAA)TCTAT site in the -59.5 bp position relative to the start site of transcription. Mutation in the ssrI gene encoding 6S RNA did not disturb P9mcc)-lac expression; these results indicate that 6S RNA does not participate in the regulation of microcin C51 operon expression.

Bacteriocins↗

PubMed2004

[Distribution of transposons Tn5044 and Tn5070 with unusual mer operons in environmental bacterial populations].

The distribution of unusual mercury resistance transposons, Tn5044 and Tn5070, was examined. A characteristic feature of Tn5044 is temperature sensitivity of its mercury operon and the presence in the mer operon of the gene homologous to RNA polymerase a subunit. Structural organization of mercury operon Tn5070, containing minimum gene set (merRTPA), differs from mer operons of both Gram-negative and Gram-positive bacteria. None of more than two thousand environmental bacterial strains displaying mercury resistance and isolated from the samples selected from different geographical regions hybridized to Tn5040- and Tn5070-specific probes. A concept on the existence of cosmopolite, endemic, and rare transposons in environmental bacterial populations was formulated.

DNA Transposable Elements↗

PubMed2005

[Activation of the expression of the microcin C51 operon upon glucose starvation of cells at the exponential growth phase].

It was earlier shown that expression of the microcin C51 operon in Escherichia coli cells is activated upon decelerated growth of cells during their transition to the stationary growth phase and depends on the sigmaS subunit of RNA polymerase. Using a single-copy construct containing the cloned promoter region of the microcin C51 operon and a promoterless lac operon (P(mcc)-lac), it was shown that the promoter of the microcin operon was also induced by stress caused by the transition of cells at the exponential growth phase into the medium without glucose as a sole carbon source. Activation of P(mcc)-lac expression upon severe glucose starvation occurred in rpoS+ and rpoS- strains. In cells carrying the rpoD800 mutation that renders the sigma70 subunit of RNA polymerase temperature-sensitive, an activation of P(mcc)-lac expression was observed at nonpermissive temperature, in contrast to its complete inhibition in E. coli cells at the phase of delayed growth. Other stressors-nitrogen starvation, high temperatures, osmotic shock, tetracycline and chloramphenicol-did not activate P(mcc)-lac expression in cells at the exponential growth phase.

Bacteriocins↗

PubMed2005

Bistability and hysteresis in epigenetic regulation of the lactose operon. Since Delbrück, a long series of ignored models.

Bistability is the capacity of a system to switch in an "all-or-none" manner between alternative steady states. This powerful concept originates from the analysis of non-linear equations driving open systems. It is one of the various patterns of regulation associated with a particular class of dynamic structures that Glansdorff and Prigogine baptised "dissipative structures". The idea of discontinuous transitions between alternative states was first formulated much earlier, by Delbrück, in 1949. Cohn and Horibata and Novick and Weiner confirmed that such transitions occur in experiments on the lactose operon carried out ten years later. Modelling with non-linear differential equations made it possible to simulate the dynamic behaviour of the lac operon, and modelling by asynchronous logical analysis elucidated the determinant role played by positive feedback circuits in the emergence of multistationarity. Nevertheless, these studies were largely ignored until the recent demonstration of the hysteretic nature of the bistable transition between alternative states of the lac operon. As originally suggested by Delbrück, the pattern of lactose consumption adopted by the bacterium is controlled epigenetically rather than genetically: the true key determinant is the direction of change of an environmental variable with respect to the structural components of the operon.

Animals↗

PubMed1987

Nucleotide sequence, promoter analysis, and linkage mapping of the unusually organized operon encoding ribosomal proteins S7 and S12 in maize chloroplast.

The nucleotide sequence of the operon encoding maize chloroplast ribosomal protein genes S7 and S12 and the promoter activity of a chimeric construct of the -10/-35 sequence of this operon (attached to a promoterless chloramphenicol acetyltransferase gene) have been determined. This operon occurs in the chloroplast genome divided in two parts: part A contains exon 1 of rpS12 (encoding the N-terminal 38 amino acid residues), whereas part B has the following structure: promoter-rpS12 (exon 2 + intron + exon 3)-spacer-rpS7-terminator. Part A is located at the approximate coordinate position 41000, whereas two copies of part B are located at two distant locations in the genome at coordinate positions 18700 and 120200. This unusual organization of the S12 operon in maize (a monocot plant) is similar to that reported in a dicot and a lower plant. The deduced amino acid sequence of maize chloroplast S7 shows 43, 38, 71, and 85% and of S12 shows 66, 72, 91 and 90% sequence identity to the corresponding sequences of Escherichia coli, Euglena gracilis, Marchantia polymorpha, and Nicotiana tabacum, respectively. The promoter upstream of rpS12 (part B) is transcriptionally active in E. coli.

Amino Acid Sequence↗

PubMed1985

[Effect of mutation changes in RNA-polymerase and transcription termination factor rho on expression of various operons in E. coli].

Six mutations, impairing DNA polymerase of E. coli in combination with the wild type gene for rho factor or ts-mutation rho 15 have been studied in relation to the expression of seven operons having different types of regulation. The expression of genes for glucose-6-phosphate dehydrogenase and 6-phosphogluconate dehydrogenase is shown to be constitutive and resistant to mutationally altered RNA polymerase and rho factor. The expression of genes for adenine phosphoribosyltransferase and of deo operon is regulated by rho dependent attenuators with attenuation being lifted incomplete medium. Mutation rho 15 decreases the level of enzymes of thr and lac operons independent of mRNA levels of these operons. Mutation rho 15 effect on posttranscriptional level is modified by mutations damaging RNA polymerase. The data obtained suppose RNA polymerase to affect all stages of realization of genetic information, beginning with promoter recognition and RNA synthesis and including the protein synthesis on mRNA.

DNA Restriction Enzymes↗

PubMed1988

Expression of the biotin biosynthetic operon of Escherichia coli is regulated by the rate of protein biotination.

In Escherichia coli biotin biosynthesis is repressed by high concentrations of exogenous biotin. This paper reports that upon high level production of the apo form of a biotinated protein, biotin operon expression was derepressed by 8-10-fold. The biotinated protein studied was the 1.3 S subunit of Propionibacterium shermanii, and transcarboxylase derepression was assayed by beta-galactosidase production in strains which carry a lacZ gene altered such that it is transcribed from biotin operon promoters. Depression of beta-galactosidase synthesis upon production of the apo 1.3 S protein was observed over a several hundred-fold range of biotin concentrations and also resulted in an increased level of biotin operon expression at maximally repressing biotin concentrations. Biotin operon derepression by apobiotin protein production seems a direct consequence of the properties of the biotin repressor protein which also functions as the ligase catalyzing the covalent attachment of biotin to apoproteins.

Bacterial Proteins↗

PubMed1985

[Riboflavin operon in Bacillus subtilis contains additional promoters].

Using the methods of molecular cloning permitted to show that riboflavin operon of Bacillus subtilis contains four promoters. Three of them are functionally active in the Bacillus subtilis system. The main promoter of the operon with regulatory region was cloned in plasmid pPL603. Cells containing the constructed plasmid pGM32 are resistant to chloramphenicol. The level of resistance is regulated by concentration of riboflavin (the effector of operon). The following model of rib-operon has been proposed: (Formula: see text).

Bacillus subtilis↗

PubMedDate not supplied

[Dependence of the activity of phi X 174 B-promoter in the expression of Escherichia coli gal-operon on the number of its copies and their orientation].

Promoter activities of different restriction fragments of the R8 DNA region of phage phi X 174 were compared. The studied DNA fragments included HindII fragment R8 (B-promoter), its left portion 49 nucleotide long, and the central segment containing 113 nucleotides generated by AluI. The promoter activity of these fragments was quantitated by the appearance of uridyltransferase and galactokinase activities in Escherichia coli clones carrying plasmids pHD68-17. The gal-promoters of these plasmids was substituted for the three aforementioned restriction fragments. The R8 region and its central part (BII-promoter) had comparable promoter activities while the left part containing the putative BI-promoter, did not induce clones with the expressed gal-operon. Clones containing 1, 2, 3 copies of the promoter fragment R8 were selected. No clones were revealed with more copies. All selected di- and tri-promoter clusters in plasmids had the same correct orientation of all inserted promoters with respect to the gal-operon. The expression of the gal-operon in E. coli was nearly directly proportional to the number of the phi X 174 B-promoters inserted before the operon.

Bacteriophage phi X 174↗

PubMed1981

[Dependence of threonine operon expression on the relA gene allelic state and the level of guanosine tetraphosphate in E. coli].

The expression of threonine operon in Rel+ and Rel- E. coli cells was studied at the transcriptional level. The rel+ genotype and the lack of a specific amino acid--threonine--are necessary for the stimulation of threonine operon transcription. Under these conditions the RNA fraction, which is thr-mRNA, is increased 2-fold. The lack of arginine or histidine in Rel+ strain does not lead to derepression of the threonine operon. It is shown that in a cell-free system 0.1 mM ppGpp stimulate the synthesis of thr-mRNA on phage DNA lambda dthr and plasmid DNA PYN1107, containing total threonine operon 1.5--2-fold. It is assumed that ppGpp stimulates the initiation of transcription. Studies on the strain carrying spoT- mutation, which significantly lowers the rate of ppGpp degradation and results in suppression of rel- phenotype, revealed positive correlation between the intracellular level of ppGpp and the thr-mRNA fraction in the total transcript.

Alleles↗

PubMed1981

[Mutation affecting the promotor region of the Escherichia coli K-12 deo-operon].

It was shown that the strain SA1030 (Das et al., 1976) contains a mutation (deoPx), which decreases deoR regulated expression of the deo operon. Thymine dependent strain harbouring deoPx mutation decreases requirement in thymine up to 8-10 microgram/ml. The deoPx mutation causes 9 fold and 3 fold decrease in the activity of thymidine phosphorylase and purine nucleoside phosphorylase, respectively in cytR-deoR- strains, i.e. under the conditions of maximal derepression of the operon. The presence of cya mutation in deoPx background causes reduction of the activity of both enzymes to basal level, no matter whether cytR or deoR repressor proteins are present or not. It is supposed that deoPx mutation blocks the activity of the deoP promoter, while the cytP promoter remains unchanged. On the other hand, the deoR mutation gives rise to a rather high level of the activity of deo enzymes in cya+cytR+ strains harbouring deoPx mutation, as compared to those found in the corresponding deoR+ strains. These data may be explained by the conception that two repressor proteins function in a cooperation with respect to repression of the deo-genes (Hammer-Jespersen, Munch-Petersen, 1975). The deoPx mutation is cotransducible (approximately 50%) with the thr gene and is located near the deo operon to the left of dra, indicating the order of markers on the chromosome to be deoPx--dra--thr. In transduction and conjugation matings deoPx mutation is characterized by a very low frequence or the absence of integration into the chromosomes of some recipient strains. These data, suggest that deoPx mutation is the result of a rearrangement of genetic material in the promoter region of the deo operon.

Enzyme Activation↗

PubMed1982

Synthesis of mRNA of malB operons at specific stages in the cell cycle of Escherichia coli.

The mRNA synthesis of the malB operons was examined using synchronous cultures of Escherichia coli. mRNA of the malB operons were synthetized in cell cycle-specific manners different from the bulk mRNA synthesis. The synthesis occurred in two stages during a cell cycle, one in the middle of the cycle and the other at the time slightly before cell division. Identification of the species of mRNA revealed that the malK-lamB operon was preferably transcribed in the former stage of the cycle while a major fraction of mRNA of the malE-malF operon was synthesized in the later stage.

Cell Cycle↗

PubMed1982

Structural and functional analysis of an rRNA operon and its flanking tRNA genes from Zea mays chloroplasts.

The complete analysis of an rRNA operon from Zea mays chloroplasts and its comparison with other plastidic or bacterial rRNA operons is presented. The maize operon contains structural genes for 16S, 23S and 4.5S rRNA species, a leader region proximal to the 16S rRNA gene and a 2.4 kb spacer between the 16S and 23S rRNA genes. Within the spacer DNA sequence are two tRNA genes which code for a tRNAIle and tRNAAla species but each gene is split by large intervening sequences of 949 and 806 bp, respectively. 4.5S rRNA is a structural equivalent of the 3' terminal region of bacterial 23S rRNA. The operon is flanked at its 5' side by a tRNAVal gene and at its 3' side by a 5S rRNA gene. Both these genes are probably not included in the large, primary precursor rRNA transcript.

Base Sequence↗

PubMed1996

Transcription termination within the regulatory nifLA operon of Klebsiella pneumoniae.

The effect of premature stop codons in the nifL gene on the expression of nifA-lacZ operon and protein fusions in Klebsiella pneumoniae was analysed in detail. Our results revealed transcriptional polarity in this operon. By dissecting the operon, intragenic regions containing Rho-dependent transcription terminators have been identified. As shown for other Rho-dependent terminators, their cytosine content is much higher than the incidence of guanines. However, other regions of the operon that have this feature did not show termination activity, suggesting that, contrary to previous reports, a correlation between these parameters cannot readily be established. Some of our results alos suggested that, in addition to polarity, other mechanisms may prevent expression of nifA when translation of nifL is altered. Their importance for efficient regulation of nitrogen fixation genes is discussed.

Bacterial Proteins↗

PubMed1996

Regulatory region with putA gene of proline dehydrogenase that links to the lum and the lux operons in Photobacterium leiognathi.

Nucleotide sequence of regulatory region (R & R) with putA gene (EMBL Accession No. U39227) from Photobacterium leiognathi PL741 has been determined, and the putA gene encoded amino acid sequence of proline dehydrogenase is deduced. Alignment and comparison of proline dehydrogenase of P. leiognathi with the proline dehydrogenase domain in the PutA protein of Escherichia coli and Salmonella typhimurium show that they are homologous. Nucleotide sequence reveals that regulatory region with the putA gene is linked to the lum and lux operons in genome; the gene order is <--putA--R & R(I)<--ter-lumQ-lumP-R & R-luxC-luxD-luxA-luxB-luxE--> (R & R: regulatory region; ter:transcriptional terminator), whereas the R & R is the regulatory region for the lum and the lux operons, ter is the transcriptional terminator for the lum operon, and R & R(I) apparently is the regulatory region for the putA and related genes. Nucleotide sequence analysis illustrates the specific inverted repeat (SIR), cAMP-CRP consensus sequence, canonical -10/-35 promoter, putative operator and Shine-Dalgarno (SD) sequence on the regulatory region R & R(I) for the putA and related genes; it suggests that the putA and related genes are simply linked to the lum and the lux operons in genome, the regulatory region R & R(I) is independent for the putA and related genes.

Amino Acid Sequence↗