Tag der mündlichen Prüfung am: The voyage of discovery is not in seeking new landscapes but in having new eyes-Marcel Proust ABBREVIATIONS 6 Abbreviations ADAM 1-aminoadamantam ADP Adenosine diphosphate ATP Adenosine triphosphate AHT Anhydrotetracycline Amp r Resistance towards ampicilin aphA3, Kan r Resistance towards kanamycin APS Ammoniumperoxodisulfate bp Base pairs BCA Bicinchoninic acid c...

In situ PCR is a technique that allows specific nucleic acid sequences to be detected in individual cells and tissues. In situ PCR and IS-RT-PCR are elegant techniques that can increase both sensitivity and throughput, but they are, at best, only semiquantitative; therefore, it is desirable first to ascertain the expression pattern by conventional means to establish the suitable conditions for ...

Numerous non-isotopic techniques have been developed in the last decade to detect and quantitate hybridization products (1). Since the polymerase chain reaction (PCR) was introduced this technique has become increasingly important for the diagnosis of bacterial and viral infections. The need for simple methods allowing quantitative evaluation of PCR products has resulted in the development of a...

Pulmonary surfactant exists in the alveolus in several distinct subtypes that differ in their morphology, composition, and surface activity. Experiments by others have implicated a serine hydrolase in the production of the inactive small vesicular subtype of surfactant (N. J. Gross and R. M. Schultz. Biochim. Biophys. Acta 1044: 222-230, 1990). Our laboratory recently identified this enzyme in ...

BACKGROUND Simultaneous and rapid detection of multiple foodborne bacterial pathogens is important for the prevention of foodborne illnesses. OBJECTIVE The aim of this study was to evaluate the use of 16S rDNA and 23S rDNA sequences as targets for simultaneous detection of eight foodborne bacterial pathogens. METHODS Nineteen bacterial oligonucleotide probes were synthesized and applied to ...

Clark, Howard, Lennell Allen, Erin Collins, Frederick Barr, Leland Dobbs, Gunther Putz, Jon Goerke, and Samuel Hawgood. Localization of a candidate surfactant convertase to type II cells, macrophages, and surfactant subfractions. Am. J. Physiol. 276 (Lung Cell. Mol. Physiol. 20): L452–L458, 1999.—Pulmonary surfactant exists in the alveolus in several distinct subtypes that differ in their morph...

We previously conducted basic research to quantify in situ hybridization (ISH) signals in rat testes. In this experimental model, we selected ribosomal RNA (rRNA) as the hybridizable RNA in paraffin sections, since it allowed us to easily analyze ISH signals expressed with digoxygenin (DIG)-labeled probes quantitatively through "posterization" of the images. We applied this method to analyze th...

We describe a fast light microscopic procedure for the simultaneous enzyme cytochemical detection of three different DNA target sequences in contrasting colors in both interphase and metaphase cell preparations. Chromosome-specific DNA probes labeled with either biotin, digoxygenin, or fluorescein were hybridized as a mixture and detected clearly and accurately by precipitates of the horseradis...

We have developed a simple and sensitive method for the rapid quantitation of mRNA from cell cultures and small tissue samples. The method combines the high sensitivity and specificity of the ribonuclease protection assay with simple handling and rapid execution of dot blotting. The use of digoxygenin-labeled cRNA probes eliminates all problems associated with radioisotopes commonly used in the...

None of the available antiretroviral drugs that are currently used in the clinic to treat infection with HIV-1 is directed against the RNase H active site of the reverse transcriptase. Here we developed a nonradioactive, 96-well plate assay designed to be used for high-throughput screening of compounds capable of inhibiting the RNase H activity of HIV-1 reverse transcriptase. We employed a tRNA...