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Nuclear run-on transcription from primary embryonic lens tissue
P S Zelenka1, L A Pallansch, M Vatal
1Laboratory of Molecular and Developmental Biology, National Eye Institute, Bethesda, Maryland 20892.
Abstract:
We have devised an in vitro RNA elongation assay (nuclear "run-on" transcription) that is suitable for use with small amounts of primary embryonic tissue. The assay is sensitive enough to detect transcription of single-copy genes in 8 X 10(5) nuclei isolated from embryonic chicken lens epithelia, and gives no detectable hybridization to unrelated DNAs, such as phi X or pBR322. We have used this assay to examine transcription of delta-crystallin and six proto-oncogenes in lens epithelia of 6-day-old embryonic chickens. The results indicate that delta-crystallin, c-myc, p53, and c-fos are actively transcribed in these cells, while c-myb, N-ras, and c-mil are not transcribed at detectable levels.
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