Protocols on Regulation of Gene Expression

Abstract

The protocols described help studying the expression of specific genes with the equipment and expertise that is usually available to most research laboratories. They are specifically intended for bacteria grown in pure cultures. The first two protocols are useful to analyse gene expression in vivo and rely on the use fusions to reporter genes such as lacZ and gfp. The third protocol requires purification of total RNA from cells and is based on the transformation of the RNA to a complementary DNA, which is then quantified by a real-time polymerase chain reaction (RT-PCR). It therefore serves to measure the abundance of specific RNAs, or changes in the levels of particular RNAs, under two different conditions. The methods described can answer different questions on the expression of a given gene and therefore complement each other.