BglII 5’...A_GATCT...3’ (5’overhang) BamHI 5’...G_GATCC...3’ (5’overhang); notice: MboI and Sau3AI = 5’..._GATC...3’ EcoRI 5’...G_AATTC...3’ (5’ overhang) HpaII 5’...C_CGG...3’ (5’ overhang); notice: CpG methylation blocked HindIII 5’...A_AGCTT...3’ (5’ overhang) KpnI 5’...GGTAC_C...3’ (3’ overhang) NcoI 5’...C_CATGG...3’ (5’ overhang); notice: Kozak sequence (RCC)ACCATGG NotI 5’...GC_GGCCGC......

In vitro recombinant DNA techniques were used to construct two new cloning vehicles, pBR324 and pBR235. These vectors, derived from plasmid pBR322, are relaxed replicating elements. Plasmid pBR324 carries the genes from pBR322 coding for resistance to the antibiotics ampicillin (Apr) and tetracycline (Tcr) and the colicin E1 structural and immunity genes derived from plasmid pMBI. Plasmid pBR32...

The Escherichia coli uvrB gene, located on a 1.5-megadalton EcoRI (fragment F, derived from transducing phage lambda b2att2 [lambda b2cI857intam6 delta (bioAB)bio-FCD+uvrB+], has been cloned in the unique EcoRI site of several "relaxed" plasmids, i.e., pMB9, pBR322, and pBH20 (= ;BR322, including the lac regulatory elements [K. Itakura, T. Hirose, R. Crea, A. D. Riggs, H. L. Heyneker, F. Boliva...

The 14 Hind111 cleavage sites on P22 DNA have been mapped. Hind111 cleavage sites were located relative to EcoRI sites by determining the molecular weights and map order of fragments produced by HindIII, or Hind111 and EcoRI digestion. Molecular weights were estimated from the electrophoretic mobility of fragments. The Hind111 fragment order was established by Hind111 cleavage of segments of th...

Polymorphisms. PstI, HindIII, and TuqI restriction fragment length polymorphisms (RFLP) in the bovine Uroplukin IB (UPKlB) gene. Source and Description ofProbe. The UPKlB probe is a .5-kb PCR product encoding a 27-kDa bovine Uroplukin IB protein associated with the urothelial asymmetric unit membrane (Yu et al., 1994). The insert was radiolabeled with [ c Y ~ ~ P I ~ C T P to a specific activit...

E. coli plays an important role in maintaining intestinal physiology. However, there are pathogenic strains that cause distinct syndromes of diarrheal disease. In this study we used a collection of 28 E. coli isolated from cattle, buffaloes and chicken obtained in the same geographical area to perform a detailed analysis of the molecular epidemiology of O157 and non O157 strains by using PCR fo...

Homeobox DNA polymorphisms (restriction fragment length polymorphisms, RFLPs) were first found in 13 individuals, and then in 121 subterranean mole rats of the Spalax ehrenbergi superspecies in Israel. Spalacidae are adapted morphologically to life underground apparently since Oligocene times, 30 million years ago. The superspecies S. ehrenbergi comprises four chromosomal species (2n = 52, 54, ...