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In the last several years, the use of double-stranded DNA templates together with thermostable-polymerase PCR has essentially replaced the use single-stranded DNA templates using the thermolabile polymerase for in vitro mutagenesis. Numerous PCR methods are now available, such as overlap-extension PCR, megaprimer PCR, and inverse PCR. All these PCR methods(More)
We characterized the pyruvate carboxylase (PC) gene by PCR amplification, subcloning, and sequencing. The coding region has 19 exons and 18 introns spanning approximately 16 kb of genomic DNA. Screening both the cDNA and the gene of individuals with the simple A form of PC deficiency revealed an 1828G-->A missense mutation in 11 Ojibwa and 2 Cree patients(More)
While the presence of a lipoyl-containing protein (protein X) separate from lipoyl transacetylase in the pyruvate dehydrogenase complex (PDC) has been known for some time, until recently only the cDNA for the yeast enzyme has been cloned. We have cloned, sequenced and characterized the cDNA encoding the human protein X and localized the protein X gene to(More)
The human mitochondrial elongation factor Tu (EF-Tu) is nuclear-encoded and functions in the translational apparatus of mitochondria. The complete human EF-Tu cDNA sequence of 1677 base pairs (bp) with a 101 bp 5'-untranslated region, a 1368 bp coding region, and a 207 bp 3'-untranslated region, has been determined and updated. The predicted protein from(More)
Yeast is an ideal host system for studying exogenous eukaryotic gene expression and for studying gene structures from recombinant plasmid DNAs and yeast artificial chromosomes (YACs). Many yeast expression studies may also require the altering of the genome itself by homologous recombination. All these studies require the screening for positive(More)
Field-effect transistors made of organic single crystals are ideal for studying the charge transport characteristics of organic semiconductor materials. Their outstanding device performance, relative to that of transistors made of organic thin films, makes them also attractive candidates for electronic applications such as active matrix displays and sensor(More)
A one-step three-stage efficient PCR method for site-directed mutagenesis has been developed. This method involves one PCR reaction with no need to add more reagents during this reaction or to do a second PCR reaction. This method utilizes a three-stage PCR cycling profile, a ddNTP-blocked restriction endonuclease fragment, and Pfu DNA polymerase. This(More)
The measurement and optimization of microstructure development in organic semiconductor films is valuable because microstructure in many cases critically impacts electronic performance. We demonstrate a general method to measure microstructure thickness dependence in thin films using surface-sensitive near edge X-ray absorbance fine structure (NEXAFS)(More)
The chromosomal location of human cytochrome c oxidase (COX) subunit VIa Liver (VIa-L) isoform related sequences has been determined by a combination of in situ hybridization and analysis of human-hamster somatic cell hybrid panels. COX VIa-L related sequences were present on chromosomes 6 and 12. It has been verified that at least two COX VIa-L genes are(More)