RESUMEN
Viral genomes are protected and organized by virally encoded packaging proteins. Heterologous production of these proteins often results in formation of particles resembling the authentic viral capsid or nucleocapsid, with cellular nucleic acids packaged in place of the viral genome. Quantifying the total protein and nucleic acid content of particle preparations is a recurrent biochemical problem. We describe a method for resolving this problem, developed when characterizing particles resembling the Menangle Virus nucleocapsid. The protein content was quantified using the biuret assay, which is largely independent of amino acid composition. Bound nucleic acids were quantified by determining the phosphorus content, using inductively coupled plasma mass spectrometry. Estimates for the amount of RNA packaged within the particles were consistent with the structurally-characterized packaging mechanism. For a bacterially-produced nucleoprotein complex, phosphorus usually provides a unique elemental marker of bound nucleic acids, hence this method of analysis should be routinely applicable.
Asunto(s)
Técnicas de Química Analítica/métodos , Proteínas de la Nucleocápside/análisis , Paramyxoviridae/química , Reacción de Biuret , Escherichia coli/genética , Escherichia coli/metabolismo , Espectrometría de Masas , Ácidos Nucleicos/análisis , Ácidos Nucleicos/metabolismo , Proteínas de la Nucleocápside/aislamiento & purificación , Proteínas de la Nucleocápside/metabolismo , Proteínas de la Nucleocápside/ultraestructura , Paramyxoviridae/genética , Paramyxoviridae/metabolismo , Paramyxoviridae/ultraestructura , Fósforo/análisis , Fosforilación , Unión Proteica , Proteínas Recombinantes/análisis , Proteínas Recombinantes/aislamiento & purificación , Proteínas Recombinantes/metabolismo , Proteínas Recombinantes/ultraestructuraRESUMEN
OBJECTIVES: To determine pharmacy faculty members' and students' attitudes and perceptions regarding the use of complementary and alternative medicine (CAM). METHODS: A survey was used to obtain data on the following information concerning CAM: general attitudes, perceived barriers to use, training received and desired, personal use, and information sources used. RESULTS: In general, attitudes of pharmacy faculty members and students towards CAM were positive. Lack of evidence for CAM practices was considered the greatest barrier to the use of CAM by faculty members and students. Many CAM therapies were perceived to be effective. A high percentage of students desire more training in the areas of herbal medicine and nutritional supplements. Faculty members and students believe that CAM should be included in the curriculum. CONCLUSIONS: General attitudes towards CAM are positive among pharmacy faculty members and students, and most believe that CAM should be taught in the pharmacy curriculum. More resources and training of faculty members may be necessary.