Nuclear run-on transcription from primary embryonic lens tissue.
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.
Zelenka
P S
PS
Laboratory of Molecular and Developmental Biology, National Eye Institute, Bethesda, Maryland 20892.
Pallansch
L A
LA
Vatal
M
M
D000546:Amanitins / Q000494:pharmacology; D000818:Animals; D002467:Cell Nucleus / Q000502:physiology; D002474:Cell-Free System; D002642:Chick Embryo; D003459:Crystallins / Q000235:genetics; D005786:Gene Expression Regulation; D066298:In Vitro Techniques; D007908:Lens, Crystalline / Q000196:embryology* / Q000502:physiology; D009693:Nucleic Acid Hybridization; D011518:Proto-Oncogene Proteins / Q000235:genetics*; D011519:Proto-Oncogenes; D014158:Transcription, Genetic
D000546:Amanitins; D003459:Crystallins; D011518:Proto-Oncogene Proteins