RESUMEN
The luxCDABE operon of Photorhabdus luminescens has been cloned and engineered as an easily mobilisable cassette flanked by sites for commonly used restriction enzymes. Constitutive and promoter probe plasmids utilising the P. luminescens luxCDABE have been constructed using a number of compatible replicons and antibiotic markers. Complementary to these plasmids, a range of promoterless and constitutive luxCDABE mini-Tn5 derivatives has been constructed. The potential of coupling mini-Tn5 luxCDABE promoter probe transposons with automated luminometry and photometry to screen for mutants that exhibit growth phase variation in gene expression is demonstrated.
Asunto(s)
Elementos Transponibles de ADN , Enterobacteriaceae/genética , Regulación Bacteriana de la Expresión Génica , Genes Reporteros/genética , Operón , Plásmidos/genética , Regiones Promotoras Genéticas , Clonación Molecular , Conjugación Genética , Enterobacteriaceae/crecimiento & desarrollo , Mediciones Luminiscentes , Fotometría , Mapeo Restrictivo , Transducción de SeñalRESUMEN
The effect of temporary clipping on the arterial endothelium of rats was examined with scanning electron microscopy. The opening pressure of four Heifetz clips was modified by changing the springs. Four different clip forces (20, 35, 45, and 65 gm), four periods of clipping (10, 30, 60, and 180 minutes), and three vessel diameters (smaller than 1 mm, 1.1 to 1.3 mm, and 1.4 to 2 mm) were compared. Different grades of endothelial damage were observed. On gross examination the damage involved a detachment of endothelium and the adherence of platelets to the subendothelial tissue. The duration of clipping seemed to be of more importance than the clip force, whereas the vessel diameter had no recognizable influence.
