RAPID NONRADIOACTIVE IN-SITU HYBRIDIZATION FOR INTERLEUKIN-2 MESSENGER-RNA WITH RIBOPROBES GENERATED USING THE POLYMERASE CHAIN-REACTION

In situ hybridization is a technique with widespread application. Howe ver, its usefulness has been limited by the need for radioactive mater ials and the requirement for the DNA to be cloned onto an appropriate vector. We have utilized the polymerase chain reaction to directly inc orporate a T7 RNA polymerase promoter sequence onto the cDNA for inter leukin-2. Digoxigenin-labelled riboprobes were then synthesized using this PCR product as a template. The digoxigenin-labelled riboprobes we re then used in non-radioactive in situ hybridization to detect messen ger RNA for interleukin-2 in mitogen stimulated peripheral blood monon uclear cells. This methodology has the potential for widespread applic ation in immunology and cytokine research.