RT-PCR amplification of P450 2C6 from rat liver, using primers in opposite orientations of exon 6, resulted in PCR products containing segments of exons joined at non-consensus splice sites. Moreover, many of the PCR products identified were composed of not only a single region containing exonic segments joined at non-consensus splice sites but, instead, of several repeats of the non-canonicall...

A Type II restriction endonuclease activity free of TaqI was prepared from Thermus Aquatious YT. The fraction contains two endonucleolytlo components with apparently different specificitiesi however the major activity is sufficiently dominant to allow partial digestion analysis of the position of recognition sites. A precise determination of the location of cleavage sites in pBR322 DNA and a co...

activities of the key enzymes of gluconeogenesis in the liver and kidney of the foetal guinea pig have been followed. The foetal guinea pigs used had a gestational age of 68-72 days. Pyruvate carboxylase was present at very low activities in the kidney of the foetus and neonatal animal and also in the foetal liver until day 50, when it increased in activity to reach a value by day 60 of more th...

An inorganic pyrophosphatase was purified over 600-fold to homogeneity as judged by polyacrylamide gel electrophoresis. The enzyme is a tetramer of Mr = 84,000, has a sedimentation coefficient of 5.8S, a Stokes radius of 3.5 nm, and an isoelectric point of 5.7. Like the enzyme of Escherichia coli, the pyrophosphatase appears to be made constitutively. The pH and temperature optima are 8.3 and 8...

Thermus aquaticus contains four major fatty acids, iso-C(15) (28%), iso-C(16) (9%), normal-C(16) (13%), and iso-C(17) (48%), when grown at 70 C, as determined by gas chromatography and mass spectrometry. Small amounts of iso-C(12), normal-C(12:1), iso-C(13), normal-C(14), iso-C(14), and normal-C(15:1) were also detected. A change in growth temperature (50 to 75 C at 5-C intervals) affects a shi...

DNA binding properties of the Type 1 DNA polymerases from Thermus aquaticus (Taq, Klentaq) and Escherichia coli (Klenow) have been examined as a function of [KCl] and [MgCl(2)]. Full-length Taq and its Klentaq "large fragment" behave similarly in all assays. The two different species of polymerases bind DNA with sub-micromolar affinities in very different salt concentration ranges. Consequently...

background: taq dna polymerase is a very important enzyme for molecular biological studies such as dna amplification and dna sequencing by the pcr. it is a standard enzyme that is used in 90% of molecular biology labs today. the aim of this study was to produce taq dna polymerase enzyme in e. coli by a reliable, practical, simple and low cost method.materials and methods: in this study, the taq...

Functional large ribosomal subunits of Thermus aquaticus can be reconstituted from ribosomal proteins and either natural or in vitro transcribed 23 S and 5 S rRNA. Omission of 5 S rRNA during subunit reconstitution results in dramatic decrease of the peptidyl transferase activity of the assembled subunits. However, the presence of some ribosome-targeted antibiotics of the macrolide, ketolide or...