CLONING DIFFERENTIALLY REGULATED GENES FROM CHONDROCYTES USING AGAROSE-GEL DIFFERENTIAL DISPLAY

The technique of RNA differential display has been used extensively to clone differentially expressed genes from a wide variety of cells and tissues. Recently, a simplified method of cloning differential displa y products, separated on agarose gels, was described. Here we report a n adaption of this method, using total RNA, to clone differentially ex pressed genes. The approach is simple and rapid, and requires only sma ll quantities of;total RNA. Utilising this approach, we have cloned th ree differentially regulated genes from chondrocytes stimulated to hyp ertrophy in vitro, and confirmed their pattern of expression by Northe rn blotting. These gene fragments were sequenced and found to correspo nd to known genes, although only one has previously been isolated from chondrocytes. (C) 1998 Elsevier Science B.V.