A novel strategy for heat-mediated activation of recombinant Taq DNA polymerase is described. A serum albumin binding protein tag is used to affinity-immobilize an E. coli-expressed Taq DNA polymerase fusion protein onto a solid support coated with human serum albumin (HSA). Analysis of heat-mediated elution showed that elevated temperatures (> 70 degrees C) were required to significantly relea...

Rogers, Jennifer, and Don D. Sheriff. Role of estrogen in nitric oxideand prostaglandin-dependent modulation of vascular conductance during treadmill locomotion in rats. J Appl Physiol 97: 756– 763, 2004; 10.1152/japplphysiol.00115.2004.—Endothelial production of nitric oxide (NO) and prostaglandins (PG) may be greater in females than in males, increasing vasodilatory responses in females. Does...

We have determined the molecular genetic basis of congenital adrenal hyperplasia due to 21-hydroxylase (21-OHase) deficiency. This common disorder of cortisol biosynthesis is HLA-linked. The haplotype HLA-(A3);Bw47;DR7 is strongly associated with 21-OHase deficiency and always carries a null allele at the locus encoding the C4A (Rodgers) form of the fourth component (C4) of complement. It seeme...

We investigated the extent of functional parasympathetic and sympathetic activity to the heart at rest and during mild to heavy dynamic exercise in conscious dogs. The animals were chronically instrumented to monitor mean arterial pressure (MAP), heart rate (HR), and terminal aortic blood flow (TAQ) and trained to run on a motor-driven treadmill. MAP, HR, and TAQ were monitored at rest and duri...

The thermostable properties of the DNA polymerase activity from Thermus aquaticus (Taq) have contributed greatly to the yield, specificity, automation and utility of the polymerase chain reaction method for amplifying DNA (1). We report here the cloning and nucleotide sequence of a DNA polymerase gene from another thermophilic bacterium — Thermus flavus (Tfl). The cloning of the DNA polymerase ...

Troponin-tropomyosin interactions. Fluorescence studies of the binding of troponin, troponin T, and chymotryptic tropo-nin T fragments to specifically labeled tropomy-osin. To the Editor: Individuals heterozygous for the factor V Leiden mutation have a seven-fold higher risk to develop venous thrombosis than wild-type (WT) individuals (1). The allele frequency of factor V Leiden in Europeans is...

dna polymerase gene from thermus aquaticus strain yt1 was amplified using venttm dna po-lymerase and cloned under the control of x.pr promoter and expression was induced by a shift in tern perature. the culture was then sonicated, and after centrifugation the lysate was treated with poly ethyleneimine followed by a salting-out step. finally the protein was precipitated with ammonium sulfate and...

Staphylococcus aureus is an economically important and a major mastitis-causing pathogen that also poses food safety and antimicrobial resistance threats. Substances in mastitic milk inhibit the Taq DNA polymerase reaction (Taq PCR) making it of limited use for detecting S. aureus mastitis. In the study reported here, a set of oligonucleotide primers of 21 and 24 bases was used in Taq-PCR to am...

The human fragile-X syndrome, a major cause of inherited mental retardation, is associated with expansion of the trinucleotide repeat GGC:GCC. Repetitive sequences in DNA are subject to slippage during catalysis by DNA polymerases. We characterized the extent of slippage of synthetic GGC:GCC repeats by various DNA polymerases: Taq DNA polymerase, Klenow fragment of DNA polymerase I, DNA Sequenc...

Point mutations in somatic cells play a role in the etiology of several classes of human pathologies. Experimental procedures are required that allow the detection and quantitation of such mutations in disease-related genes in tissue biopsy samples without the need for the selection of mutated cells. We describe the genotypic analysis of single base pair mutations in the Taq I endonuclease reco...