N Curach

Prospecting for novel lipase genes using PCR

A PCR method suitable for the isolation of lipase genes directly from environmental DNA is described. The problems associated with the low levels of similarity between lipase genes were overcome by extensive analysis of conserved regions and careful primer design. Using this method, a lipase gene (oli-lipase) was isolated directly from environmental DNA. This lipase showed less than 20% similarity with other known lipases at the amino acid level. The study also revealed that distantly related members of the alpha/beta hydrolase superfamily share similar conserved motifs with the lipases, thus making these genes targets for gene prospecting by PCR.

High levels of variability in immune response using antigens from two reproductive proteins in brushtail possums

Immune-based fertility control is being considered as an effective long-term approach for controlling the pest brushtail possum (Trichosurus vulpecula) population in New Zealand. This relies heavily on the immune response of each immunised possum. A strong and lasting immune response in the majority of individuals in a population is essential. In this study, possums and the model macropod species, the tammar wallaby (Macropus eugenii) were immunised with either a luteinising hormone or androgen receptor synthetic peptide coupled to the carrier molecule KLH (keyhole limpet haemocyanin). The antibody response of wallabies to the antigens was relatively uniform. In contrast, the possum immunoglobulin response to both synthetic peptides and KLH was variable. The apparent high level of variation in the immune response of possums raises questions about the feasibility of using these two antigens to control possum numbers in New Zealand.

Overview of Gene Expression Using Filamentous Fungi

Filamentous fungi are lower eukaryotes increasingly used for expression of foreign proteins ranging from industrial enzymes originating from other fungi and bacteria to proteins of mammalian origin, such as antibodies and growth factors. Their strengths include an excellent capacity for protein secretion and their eukaryotic protein processing machinery. Proteins secreted from filamentous fungi are modified in the secretory pathway, with folding, proteolytic processing, and addition of glycans being the main modifications. Unlike from many other expression systems, however, plasmids and host strains for expression of gene products in filamentous fungi are not readily available commercially, and the expression system must thus be stitched together in the laboratory. In this overview, the key elements of fungal expression systems are discussed from a practical point of view and with a view towards the future. The principles and considerations presented here can be applied to a range of filamentous fungi.