Use of virion DNA as a cloning vector for the construction of mutant and recombinant herpesviruses.

We have developed improved procedures for the isolation of deletion mutant, point mutant, and recombinant herpesvirus saimiri. These procedures take advantage of the absence of NotI and AscI restriction enzyme sites within the viral genome and use reporter genes for the identification of recombinant viruses. Genes for secreted engineered alkaline phosphatase and green fluorescent protein were placed under simian virus 40 early promoter control and flanked by NotI and AscI restriction sites. When permissive cells were cotransfected with herpesvirus saimiri virion DNA and one of the engineered reporter genes cloned within herpesvirus saimiri sequences, recombinant viruses were readily identified and purified on the basis of expression of the reporter gene. Digestion of recombinant virion DNA with NotI or AscI was used to delete the reporter gene from the recombinant herpesvirus saimiri. Replacement of the reporter gene can be achieved by NotI or AscI digestion of virion DNA and ligation with a terminally matched fragment or, alternatively, by homologous recombination in cotransfected cells. Any gene can, in theory, be cloned directly into the virion DNA when flanked by the appropriate NotI or AscI sites. These procedures should be widely applicable in their general form to most or all herpesviruses that replicate permissively in cultured cells.

Handing Down and Adaption: The Construction of the Material World of Health Care Technology in Spain (1855-1955)

This historical study uses qualitative methods to analyze and describe the components of the material world of nursing care in Spain between 1855 and 1955 based on the analysis of eight nurse training manuals. A total of 360 objects and 45 procedures were recorded. Manual analysis was carried out concurrently with data collection based on the Grounded Theory approach. Findings show that the material world of health care was composed of objects that were handed down by the medical profession to health care professionals and adapted objects, improvised mainly out of everyday household items. While the handing down of medical tools and instruments could be said to be a theoretical and technical achievement, it is not clear whether it was also a scientific accomplishment. The improvisation of objects out of everyday household items promoted by the manuals highlights the artisan-like and ingenious nature of nursing practice, which should be explored further in future studies to provide a greater understanding and promote the recognition of these objects as a health care technology.