Analysis of Gene Regulation in Rabbit Corneal Epithelial Cells Induced by Ultraviolet Radiation

• DOI: 10.3390/ijerph2005010051
• Publication Date: 2008-10-25T17:52:15Z
• Authors: Jacqueline J. St...; Christian Rogers; Carolyn B. Howard; Caronda Moore; Lai-Man Chan
• Keywords: Differential display; Cloning (programming); Ultraviolet light

Ultraviolet (UV)-induced cataracts are becoming a major environmental health concern because of the possible decrease in stratospheric ozone layer. Experiments were designed to isolate gene(s) affected by UV irradiation rabbit cornea tissues using fluorescent differential display-reverse transcription-polymerase chain reaction (FDDRT-PCR). The epithelial cells grown standard medium for 2 or 4 hours post treatment. Cornea irradiated with UVB 20 minutes. RNA was extracted and amplified reverse transcriptase-polymerase poly A+ specific anchoring primers random arbitrary primers. Polyacrylamide gel electrophoresis revealed several differentially expressed genes untreated versus cells. Complimentary DNA (cDNA) fragments resulting from mRNAs eluted re-amplified. re-amplified PCR products cloned directly into PCR-TRAP cloning system. These data showed that FDDRT-PCR is useful technique elucidate UV-regulated gene expressions. Future experiments will involve sequence analysis inserts. identification these through could lead better understanding cataract formation via damage mechanisms prevention.