Expression, Glycosylation, and Secretion of an Aspergillus Glucoamylase by Saccharomyces cerevisiae

A strain of Saccharomyces cerevisiae capable of simultaneous hydrolysis and fermentation of highly polymerized starch oligosaccharides was constructed. The Aspergillus awamori glucoamylase enzyme, form GAI, was expressed in Saccharomyces cerevisiae by means of the promoter and termination regions from a yeast enolase gene. Yeast transformed with plasmids containing an intron-free recombinant glucoamylase gene efficiently secreted glucoamylase into the medium, permitting growth of the transformants on starch as the sole carbon source. The natural leader sequence of the precursor of glucoamylase (preglucoamylase) was processed correctly by yeast, and the secreted enzyme was glycosylated through both N- and O-linkages at levels comparable to the native Aspergillus enzyme. The data provide evidence for the utility of yeast as an organism for the production, glycosylation, and secretion of heterologous proteins.     

Caspase activation in equine influenza virus induced apoptotic cell death.

Equine influenza virus (EIV) is the leading cause of acute respiratory infection in horses worldwide. In recent years, the precise mechanism by which influenza infection kills host cells is being re-evaluated. In this report, we examined whether caspases, a group of intracellular proteases, are activated following EIV infection and contribute to EIV-mediated cell death. Western blotting analysis indicated that a nuclear target of caspase-3, poly(ADP-ribose) polymerase (PARP) was proteolytically cleaved in EIV-infected MDCK cells, but not in mock-infected cells. In comparison with caspase-3 specific inhibitor Ac-DEVD-CHO, a general caspase inhibitor Boc-D-FMK provided much stronger inhibition of EIV-induced cytopathic effect and apoptosis. Our results suggest that EIV may activate more than one caspase. Caspase activation and cleavage of its cellular targets may play a critical role in EIV-mediated cytotoxicity.     

Quantification of historical livestock importation into New Zealand 1860–1979

AIMS: To quantify the numbers of live cattle, sheep and poultry imported into New Zealand and, where possible, their country of origin from 1860 to 1979.

METHODS: Information on the origin and number of live animal importations into New Zealand was collected for cattle, sheep and poultry for the period 1868–1979 from the annual reports compiled by the New Zealand Registrar General's Office, Government Statistician's Office, Census and Statistics Office, Census and Statistics Department, Customs Department and Department of Statistics. Census data from 1851 to 1871 were also used to estimate the livestock population during this period. The number of animals imported and the mean population for each species in a decade were determined, and the major countries of origin were identified.

RESULTS: A large number of cattle (53,384) and sheep (604,525) were imported in the 1860s, and then there was a marked reduction in importations. Live poultry were imported in relatively small numbers (20,701) from 1880 to 1939, then 1,564,330 live poultry were imported between 1960 and 1979. Australia was the predominant country of origin for sheep between 1868 and 1959 (51,347/60,918; 84.3%) and of cattle between 1868 and 1979 (10,080/15,157; 66.5%). Only 6,712 (11.0%) sheep and 3,909 (25.8%) cattle were imported from the United Kingdom over the same periods, and even fewer from other countries.

CONCLUSIONS: The collated data and historical reports show that from 1860 to 1979 Australia has been the main source of livestock introduced into New Zealand. The pattern of importation showed that large numbers of cattle and sheep were initially imported in the 1860s, probably in response to rapid agricultural expansion. Thereafter importations continued at much reduced numbers. In contrast, relatively small numbers of poultry were introduced until the 1960s when large numbers were imported as part of the development of a modern high-production industry. The overall pattern for both cattle and sheep was of a bottleneck event, as initially a relatively limited number of animals arrived from outside populations, followed by population expansion with ongoing but limited immigration (admixture). Investigation into the genetic population structure of New Zealand's cattle and sheep, as well as their host-associated microorganisms, could reflect the impact of these early historical events.