Rat Dgka (Diacylglycerol kinase alpha) ELISA Kit

This study investigated the production of triacylglycerols in cells of the wild type of Escherichia coli and of a strain with a deleted diacylglycerol kinase gene (dgkA). By overexpression of atfA from Acinetobacter baylyi ADP1 and fadD from E. coli in the dgkA deletion mutant, cellular contents of up to 4.9%(w/w) triacylglycerols could be achieved in batch cultivation. Furthermore, heterologous expression of atfA relieves the negative effects of dgkA deletion on growth. Process optimization and fed-batch fermentation resulted in the production of 530 mg l(-1) triacylglycerols and a maximal content of 8.5 % (w/w) triacylglycerols of the cell dry mass. This clearly exceeded all previous results concerning triacylglycerol production in E. coli. Furthermore, the production of extracellular free fatty acids and fatty acid ethyl esters was investigated. Like triacylglycerols, both products are potential biofuels, and we show their continuous production in a repeated batch process, with recovery of the production cells.

Genomic amplification can lead to the activation of cellular protooncogenes during tumorigenesis, and is observed in most, if not all, human malignancies, including adenocarcinomas of lung and esophagus. Using a two-dimensional restriction landmark genomic scanning technique, we identified five NotI/HinfI fragments with increased genomic dosage in an adenocarcinoma of the gastroesophageal junction. Four of these amplified fragments were matched within three contigs of chromosome 12 using the bioinformatics tool, Virtual Genome Scan. All three of the contigs map to the 12q13-q14 region, and the regional amplification in the tumor was verified using comparative genomic hybridization analysis. The 12q14 amplicon was characterized using sequence tagged site-amplification mapping with DNA from paired normal-tumor tissues of 75 gastroesophageal and 37 lung adenocarcinomas. The amplicon spans a region of >12 Mb between genes DGKA and BLOV1. The core-amplified domain was determined to be <0.5 Mb between marker WI-12457 and gene IFNG. However, MDM2, a well-documented oncogene of the region, is outside the core-domain. Eleven genes and expressed sequence tags within the amplicon were selected for quantitative reverse transcription-PCR, and DYRK2, a member of the dual-specificity kinase family, was overexpressed in all of the tumors showing gene amplification. Among the sequence tagged site/ expressed sequence tag/gene markers tested, DYRK2 demonstrated the highest DNA copy number and the highest level of mRNA overexpression in the tumors. Moreover, DYRK2 mRNA overexpression (>2.5-fold of normal mean) was found in 18.6% of additional 86 lung adenocarcinomas in an assay using oligonucleotide microarrays. DYRK2 mRNA overexpression occurs more frequently than gene amplification in both esophageal and lung adenocarcinomas. This is the first report of amplification and overexpression of DYRK2 in any tumor type.