Proteases are widely used to remove affinity and solubility tags from recombinant proteins to avoid potential interference of these tags with the structure and function of the fusion partner. In recent years, great interest has been seen in use of the human rhinovirus 3C protease owing to its stringent sequence specificity and enhanced activity. Like other proteases, activity of the human rhinovirus 3C protease can be affected in part by the buffer components and additives that are generally employed for purification and stabilization of proteins, hence, necessitate their removal by tedious and time-consuming procedures before proteolysis can occur. To address this issue, we examined the effect of elution buffers used for common affinity based purifications, salt ions, stability/solubility and reducing agents, and detergents on the activity of the human rhinovirus 3C protease using three different fusion proteins at 4°C, a temperature of choice for purification of many proteins. The results show that the human rhinovirus 3C protease performs better at 4°C than the frequently used tobacco etch virus protease and its activity was insensitive to most of the experimental conditions tested. Though number of fusion proteins tested is limited, we expect that these finding will facilitate the use of the human rhinovirus 3C protease in recombinant protein production for pharmaceutical and biotechnological applications.
Pesticides are a large and diverse group of chemicals used to kill and eradicate rodents, fungi, insects, and weeds. The present study was designed to check the effect of pesticides on liver as well as kidney function of the workers engaged in occupations in which there is persistent exposure to pesticides, (dealers, gardeners and owners of gardens), and to check the prevalence of hepatitis in these workers indistrict Swat, Khyber Pakhtunkhwa, Pakistan. Blood samples ISSN 2157-6076 2013 www.macrothink.org/jbls 44 were collected and history was taken from different groups of individuals exposed to pesticides. Four groups were made, owners, gardeners, dealers and control, which had approximately same age and were engaged in this field for an average of 8 years. A significant decrease in hemoglobin concentration was noticed in all the exposed groups. The level of glutamate pyruvate transaminase, uric acid, bilirubin (direct and total) and calcium was high among exposed groups compared to control. The prevalence of both hepatitis B and C was also high in exposed groups. Pesticides significantly affected the hemoglobin level, and altered liver and kidney functions. The same study may be preceded to other biochemical and hematological parameters considering a large number of exposed persons.
Journal of Biology and Life Science
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