Deoxyribonucleic Acid-Ribonucleic Acid Hybridization Studies on the l -Arabinose Operon of Escherichia coli B/r

Abstract

An increase in the rate of synthesis of ara-specific messenger ribonucleic acid as measured by deoxyribonucleic acid-ribonucleic acid hybridization has been detected in the induced wild-type (ara+) strain of Escherichia coli B/r as compared with the uninduced control, thus providing evidence that regulation of the positively controlled L-arabinose operon is at the level of transcription.

Functional limits of the araIc promoter suggest an additional regulatory site for araBAD expression

“…When required, L-arabinose was added to a final concentration of 0.4% at a turbidity of 100 Klett units and the cells were grown to a turbidity of 120 Klett units before labeling was performed. RNA was isolated as described by Wilcox et al (26). DNA-RNA hybridization was performed as described by Miyada et al (17) except that 0.5 pug of the M13-mp2 and M13-mp2::araB plus-strand DNAs were used in the hybridizations.…”

Effect of araC gene product on catabolite repression in the L-arabinose regulon

“…1); therefore, isomerase production occurs only as a consequence of araC protein added to the system and not as a consequence of in vitro synthesis. The isolation of phage and preparation of deoxyribonucleic acid were according to the procedures of Wilcox et al (8) as modified by Wilcox et al (7).…”

Isolation of specialized transducing bacteriophage lambda carrying genes of the L-arabinose operon of Escherichia coli B/r

“…By directed integration of a heat-inducible, lysis-defective bacteriophage lambda (AcI857S7) into the L-arabinose region, we have been able to isolate plaqueforming ara-transducing phages (Apara), defective ara-transducing phages (Adara), and defective leucine-transducing phages (Adleu). [20][21][22][23][24][25][26][27][28] araA 766 NL20-029 araClOl NL20-030 araC12 NL [20][21][22][23][24][25][26][27][28][29][30][31][32][33][34][35][36] A(gal attA chiA bioA uvrB thi A(leu-ara) AJ_P NL 20-037 A(gal attX chiA bioA uvrB) thi araCc67 araD139 NL [20][21][22][23][24][25][26][27][28][29][30][31][32][33][34][35][36][37][38][39] araB27 NL araB24 NL araB14 NL araB43 NL araBA87 NL 20 araB25 NL araB46 NL 20-082 araC98 NL 20-083 araC97 NL araC124 This study This study B. Gielow B. Gielow B. Gielow B. Gielow This study…”

“…NL NL NL 20-804 NL NL NL 20-808 NL NL NL NL NL NL 20-815 NL NL NL NL NL NL NL NL [20][21][22][23][24][25][26][27][28][29][30][31][32][33][34][35][36][37][38] NL A(gal attX chiA bioA uvrB) thi leu (XcI857S7 integrated in leucine) NL 20-037 (polylysogenic for XcI857S7 integrated in araB) NL 20-037 (polylysogenic for AcI857S7 integrated in araB) NL 20-037 (single Aara-TRANSDUCING BACTERIOPHAGE 1045 using EMBA and EMBSA, where ara+ transductants appear as dark centers within individual plaques on a pink lawn after 36 h of incubation at 32 C.…”

“…After equilibrium centrifugation the phage bands were withdrawn by puncturing the side of the tube with a hypodermic syringe fitted with a 24-gauge needle. The preparations so obtained were dialyzed against 0.01 M Tris-glycine, 0.01 M MgSO4, pH 7.4 (for the A phage preparations), or 0.1 M NaCl, 0.1 M Na,HPO,-NaH,PO4, pH 7.1 (for the 48OiA phage preparations), and stored at 4 C. Phage deoxyribonucleic acid (DNA) was extracted from purified phage as previously described (23). The separated strands of the AcI857S7 and Aara phage DNAs were obtained as in (20).…”