Mutations in beta ` subunit of Escherichia coli RNA polymerase perturb the activator polymerase functional interaction required for promoter clearance

P>Transcription activator C employs a unique mechanism to activate mom gene of bacteriophage Mu. The activation process involves, facilitating the recruitment of RNA polymerase (RNAP) by altering the topology of the promoter and enhancing the promoter clearance by reducing the abortive transcription. To understand the basis of this multi-step activation mechanism, we investigated the nature of the physical interaction between C and RNAP during the process. A variety of assays revealed that only DNA-bound C contacts the beta' subunit of RNAP. Consistent to these results, we have also isolated RNAP mutants having mutations in the beta' subunit which were compromised in C-mediated activation. Mutant RNAPs show reduced productive transcription and increased abortive initiation specifically at the C-dependent mom promoter. Positive control (pc) mutants of C, defective in interaction with RNAP, retained the property of recruiting RNAP to the promoter but were unable to enhance promoter clearance. These results strongly suggest that the recruitment of RNAP to the mom promoter does not require physical interaction with C, whereas a contact between the beta' subunit and the activator, and the subsequent allosteric changes in the active site of the enzyme are essential for the enhancement of promoter clearance.

In vivo clearance of Japanese encephalitis virus by adoptively transferred virus specific cytotoxic T lymphocytes

Japanese encephalitis virus (JEV) is a positive stranded RNA virus that belongs to the flavivirus group, JEV infection damages the central nervous system (CNS) and is one of the main causative agents of acute encephalitis, H-2 restricted virus-specific cytotoxic T lymphocytes (CTL) have been generated specifically against JEV in our laboratory and these CTL have been shown to protect mice against lethal challenge with JEV, Virus replication was found to be inhibited in the brains of animals that mere adoptively transferred with JEV specific CTL as revealed by immunohistological staining as,veil as viral plaque assays. We further show that virus specific CTL could be recovered from such protected mice as long as 45 days after adoptive transfer.

An Optical Fiber Bundle Sensor for Tip Clearance and Tip Timing Measurements in a Turbine Rig

When it comes to measuring blade-tip clearance or blade-tip timing in turbines, reflective intensity-modulated optical fiber sensors overcome several traditional limitations of capacitive, inductive or discharging probe sensors. This paper presents the signals and results corresponding to the third stage of a multistage turbine rig, obtained from a transonic wind-tunnel test. The probe is based on a trifurcated bundle of optical fibers that is mounted on the turbine casing. To eliminate the influence of light source intensity variations and blade surface reflectivity, the sensing principle is based on the quotient of the voltages obtained from the two receiving bundle legs. A discrepancy lower than 3% with respect to a commercial sensor was observed in tip clearance measurements. Regarding tip timing measurements, the travel wave spectrum was obtained, which provides the average vibration amplitude for all blades at a particular nodal diameter. With this approach, both blade-tip timing and tip clearance measurements can be carried out simultaneously. The results obtained on the test turbine rig demonstrate the suitability and reliability of the type of sensor used, and suggest the possibility of performing these measurements in real turbines under real working conditions.

NUMERICAL STUDY OF THE 3D FLOWFIELD IN A TRANSONIC COMPRESSOR ROTOR WITH A MODELLING OF THE TIP CLEARANCE FLOW.

This paper describes a computer code aimed at solving the equations of three dimensional viscous compressible flow in turbomachine geometries. The code is applied to the study of the flowfield in a transonic axial compressor rotor at design speed at both maximum flow and towards stall. The predicted flowfield is compared with the laser measurements and the performance of the code discussed. In addition the discussion highlights the change in the predicted endwall and tip clearance flows as the rotor operating point is moved towards stall.

Experimental study of tip clearance flow in a radial inflow turbine

This paper describes an experimental investigation of tip clearance flow in a radial inflow turbine. Flow visualization and static pressure measurements were performed. These were combined with hot-wire traverses into the tip gap. The experimental data indicates that the tip clearance flow in a radial turbine can be divided into three regions. The first region is located at the rotor inlet, where the influence of relative casing motion dominates the flow over the tip. The second region is located towards midchord, where the effect of relative casing motion is weakened. Finally a third region exists in the exducer, where the effect of relative casing motion becomes small and the leakage flow resembles the tip flow behaviour in an axial turbine. Integration of the velocity profiles showed that there is little tip leakage in the first part of the rotor because of the effect of scraping. It was found that the bulk of tip leakage flow in a radial turbine passes through the exducer. The mass flow rate, measured at four chordwise positions, was compared with a standard axial turbine tip leakage model. The result revealed the need for a model suited to radial turbines. The hot-wire measurements also indicated a higher tip gap loss in the exducer of the radial turbine. This explains why the stage efficiency of a radial inflow turbine is more affected by increasing the radial clearance than by increasing the axial clearance.