Comparative analyses of ESTs and cDNAs with genomic DNA predict a high frequency of alternative splicing in human genes. However, there is an ongoing debate as to how many of these predicted splice variants are functional and how many are the result of aberrant splicing (or 'noise'). To address this question, we compared alternatively spliced cassette exons that are conserved between human and ...

Here we describe the development of a baculovirus vector expression cassette containing rearranged baculovirus-derived genetic regulatory elements. This newly designed expression cassette conferred significant production improvements to the baculovirus expression vector system (BEVS), including prolonged cell integrity after infection, improved protein integrity, and around 4-fold increase in r...

Professor Sir John Burn is Professor of Clinical Genetics at Newcastle University and a non-executive director of NHS England where at the time of submission he was deputy chair of Specialised Commissioning at NHS England. He helped write the 2003 government White Paper on Genetics which used warfarin genotyping as an example of future deployment of Pharmacogenetics. He has been working for 10 ...

We report here an alternative and easy approach to generate sufficient quantity of minimal gene expression cassette (promoter–open reading frame–terminator) DNA by PCR amplification using high fidelity DNA polymerases. Isolating the minimal gene cassette DNA in large quantities by restriction digestion and gel extraction for biolistic-mediated ‘clean DNA’ transformation, is not only a cumbersom...

Fast onset and high-level neurospecific transgene expression in vivo is of importance for many areas in neuroscience, from basic to translational, and can significantly reduce the amount of vector load required to maintain transgene expression in vivo. In this study, we tested various cis elements to optimize transgene expression at transcriptional, posttranscriptional, and posttranslational le...

The aim of this work was the construction of a cassette, i.e., a non-replicative molecule formed by linkage of an antibiotic resistance gene and a multiple cloning site. This cassette would allow the cloning and analysis of a wide range of replicons. The aac(6')-lc amikacin gene was isolated and ligated to the multiple cloning site of the pUC18 vector. This construction was HindIII digested and...

40 Fast onset and high level neuro-specific transgene expression in vivo is of importance for many 41 areas in neuroscience, from basic to translational, and can significantly reduce the amount of 42 vector load required to maintain transgene expression in vivo. In this study, we tested various cis 43 elements to optimize transgene expression at transcriptional, post-transcriptional and post44 ...

We developed a simple method of generating markerless deletions in the Escherichia coli chromosome. The method consists of two recombination events stimulated by lambda Red recombinase. The first recombination replaced a target region with a marker cassette and the second then eliminated the marker cassette. The marker cassette included an antibiotic resistant gene and a negative selection mark...