Transcription of the chicken alpha 2 (Type I) collagen gene by homologous cell-free extracts.

摘要: We have used two methods to detect specific transcription of the chicken alpha 2 (type I) collagen gene in cell-free extracts derived from Rous sarcoma virus-transformed embryo fibroblasts. The first method is a modification S1 nuclease mapping procedure which utilizes DNA probe labeled with 32P at 5' end HindIII linker originally clone promoter region into PBR322. distinguishes newly made, RNA endogenous and nonspecific transcripts. Using this we found that whole cell support accurate initiation template. Addition either creatine phosphate, GTP, or UTP concentrations approximately 3 5 mM was stimulate polymerase II by 5- 10-fold. second employs an avian myeloblastosis virus reverse transcriptase-catalyzed primary extension procedure, rendered vitro-specific use pBR322 fragment as primer. These techniques should be useful for analyzing other types extracts.