Expression Vectors Used in Project-Oriented Teaching Laboratories

 

Susan J. Karcher, David J. Asai, and Chris J. Staiger

Tested Studies in Laboratory Teaching, 2004, Volume 25

Abstract

This chapter describes a multi-week project taught in an upper level cell and molecular biology laboratory. Isolation of sufficient quantities of rare proteins from biological tissues can be difficult. These native proteins are often required for raising antibodies, and for studying enzymatic or regulatory function. Expression vectors are used to obtain large quantities of proteins. The cDNA encoding a protein is cloned into an expression vector. The fusion protein is over-expressed in E. coli, and large quantities of the recombinant protein are obtained by affinity chromatography. SDS-polyacrylamide gel electrophoresis is used to determine the size of the proteins isolated.

Keywords:  E. coli, SDS-PAGE, protein isolation, expression vector, project-oriented laboratory

University of Nevada, Las Vegas (2003)