DNA sequence divergence among derivatives of Escherichia coli K-12 detected by arbitrary primer PCR (random amplified polymorphic DNA) fingerprinting.

Derivatives of Escherichia coli K-12 of known ancestry were characterized by random amplified polymorphic DNA (RAPD) fingerprinting to better understand genome evolution in this family of closely related strains. This sensitive method entails PCR amplification with arbitrary primers at low stringency and yields arrays of anonymous DNA fragments that are strain specific. Among 150 fragments scored, eight were polymorphic in that they were produced from some but not all strains. Seven polymorphic bands were chromosomal, and one was from the F-factor plasmid. Five of the six mapped polymorphic chromosomal bands came from just 7% of the genome, a 340-kb segment that includes the terminus of replication. Two of these were from the cryptic Rac prophage, and the inability to amplify them from strains was attributable to deletion (excision) or to rearrangement of Rac. Two other terminus-region segments that resulted in polymorphic bands appeared to have sustained point mutations that affected the ability to amplify them. Control experiments showed that RAPD bands from the 340-kb terminus-region segment and also from two plasmids (P1 and F) were represented in approximate proportion to their size. Optimization experiments showed that the concentration of thermostable polymerase strongly affected the arrays of RAPD products obtained. Comparison of RAPD polymorphisms and positions of strains exhibiting them in the pedigree suggests that many sequence changes occurred in these historic E. coli strains during their storage. We propose that the clustering of such mutations near the terminus reflects errors during completion of chromosome replication, possibly during slow growth in the stab cultures that were often used to store E. coli strains in the early years of bacterial genetics

Xenopus H-RasV12 promotes entry into meiotic M phase and cdc2 activation independently of Mos and p42MAPK

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Analysis of CRP-CytR interactions at the Escherichia coli udp promoter.

Multiprotein complexes regulate the transcription of certain bacterial genes in a sensitive, physiologically responsive manner. In particular, the transcription of genes needed for utilization of nucleosides in Escherichia coli is regulated by a repressor protein, CytR, in concert with the cyclic AMP (cAMP) activated form of cAMP receptor protein (CRP). We studied this regulation by selecting and characterizing spontaneous constitutive mutations in the promoter of the udp (uridine phosphorylase) gene, one of the genes most strongly regulated by CytR. We found deletions, duplications, and point mutations that affect key regulatory sites in the udp promoter, insertion sequence element insertions that activated cryptic internal promoters or provided new promoters, and large duplications that may have increased expression by udp gene amplification. Unusual duplications and deletions that resulted in constitutive udp expression that depended on the presence of CytR were also found. Our results support the model in which repression normally involves the binding of CytR to cAMP-CRP to form a complex which binds to specific sites in the udp promoter, without direct interaction between CytR protein and a specific operator DNA sequence, and in which induction by specific inducer cytidine involves dissociation of CytR from cAMP-CRP and the RNA polymerase interaction with cAMP-CRP bound to a site upstream of then transcription start point. The stimulation of udp expression by CytR in certain mutants may reflect its stabilization of cAMP-CRP binding to target DNA and illustrates that only modest evolutionary changes could allow particular multiprotein complexes to serve as either repressors or transcriptional activators

Raman spectroscopy: comparing the “fingerprints” of C6 glioma and mesenchymal stem cells

peer reviewedUsing Coherent anti-Stokes Raman scattering (CARS) mapping we compared the Raman spectra of rat’s C6 glioma cells with those of mesenchymal stem cells. Raman spectroscopy revealed a striking similarity of scattering spectra from rat’s mesenchymal stem cells and C6 glioma cells. Our results indicate possible relations between the Mesenchymal Stem Cells (MSCs) and C6 glioma cells, which is in accordance with the studies reporting expression of common antigens in stem cells and various types of tumo

Excitonic photoluminescence quenching by impact ionization of excitons and donors in GaAs/Al(0.35)Ga(0.65)As quantum wells with an in-plane electric field

We present a detailed experimental study on photoluminescence quenching due to exciton and donor impact ionization by accelerated electrons under an in-plane nanosecond duration electric field created in GaAs/Al(0.35)Ga(0.65)As quantum wells. From the photoluminescence transients measured by the time-correlated single-photon counting technique, we have determined the experimental conditions under which donor impact ionization can have an influence on quenching of the excitonic photoluminescence. The coefficient of twodimensional exciton impact ionization has been estimated; its dependences on the applied electric field, lattice temperature, and width of the quantum wells are given

cysQ, a gene needed for cysteine synthesis in Escherichia coli K-12 only during aerobic growth.

The initial steps in assimilation of sulfate during cysteine biosynthesis entail sulfate uptake and sulfate activation by formation of adenosine 5'-phosphosulfate, conversion to 3'-phosphoadenosine 5'-phosphosulfate, and reduction to sulfite. Mutations in a previously uncharacterized Escherichia coli gene, cysQ, which resulted in a requirement for sulfite or cysteine, were obtained by in vivo insertion of transposons Tn5tac1 and Tn5supF and by in vitro insertion of resistance gene cassettes. cysQ is at chromosomal position 95.7 min (kb 4517 to 4518) and is transcribed divergently from the adjacent cpdB gene. A Tn5tac1 insertion just inside the 3' end of cysQ, with its isopropyl-beta-D-thiogalactopyranoside-inducible tac promoter pointed toward the cysQ promoter, resulted in auxotrophy only when isopropyl-beta-D-thiogalactopyranoside was present; this conditional phenotype was ascribed to collision between converging RNA polymerases or interaction between complementary antisense and cysQ mRNAs. The auxotrophy caused by cysQ null mutations was leaky in some but not all E. coli strains and could be compensated by mutations in unlinked genes. cysQ mutants were prototrophic during anaerobic growth. Mutations in cysQ did not affect the rate of sulfate uptake or the activities of ATP sulfurylase and its protein activator, which together catalyze adenosine 5'-phosphosulfate synthesis. Some mutations that compensated for cysQ null alleles resulted in sulfate transport defects. cysQ is identical to a gene called amtA, which had been thought to be needed for ammonium transport. Computer analyses, detailed elsewhere, revealed significant amino acid sequence homology between cysQ and suhB of E. coli and the gene for mammalian inositol monophosphatase. Previous work had suggested that 3'-phosphoadenoside 5'-phosphosulfate is toxic if allowed to accumulate, and we propose that CysQ helps control the pool of 3'-phosphoadenoside 5'-phosphosulfate, or its use in sulfite synthesis

Microwave sensor based on modulation-doped GaAs/AlGaAs structure

We propose a microwave diode based on a modulation-doped GaAs/Al(0.25)Ga(0.75)As structure. The principle of the diode operation relies on a non-uniform heating of the two-dimensional electron gas in microwave electric fields arising due to the asymmetric shape of the device. The voltage sensitivity of the diode at room temperature is dose to 0.3 V W(exp -1) at 10 GHz, which is comparable to the value obtained using similarly shaped and sized diodes based on bulk n-GaAs. At liquid nitrogen temperature, the voltage sensitivity strongly increases reaching a value of 20 V W(exp -1) due to the high mobility of the two-dimensional electron gas. The detected signal depends linearly on power over 20 dB, until hot-electron real-space-transfer effects begin to predominate. We discuss noise temperature measurements at 10 GHz, consider the frequency dependence of the voltage sensitivity in the microwave range and compare the performance data of the proposed device and the asymmetrically shaped bulk GaAs diode within the 10 GHz-2.5 THz frequency range