Development of a Cloning Strategy to Enable the Isolation of Novel Membrane Proteins

[摘要] The aim of the project was to develop a cloning strategy for the isolation of novel membrane proteins. The scheme evolved from the surface antigen screening system developed by Seed and Aruffo (1987). The system was adapted to enable the cloning of cDNAs coding for membrane proteins to which no antibodies exist. This was achieved by taking a previously cloned membrane protein gene (CD2), deleting its transmembrane (Tm) and cytosolic regions, and replacing them with the polylinker and stuffer sequences of CDM8. cDNAs were then sub-cloned into this expression vector to generate a fusion library. which was then screened for surface expression of the CD2 protein.

[发布机构] University:University of Glasgow

[关键词] Genetics, Molecular biology