DNA-dependent transcription of adenovirus genes in a soluble whole-cell extract
TLDR
A cell-free system for studying the synthesis of mRNA in mammalian cells, which consists of a dialyzed and concentrated whole-cell extract derived from HeLa cells, small molecules and cofactors needed for transcription, and exogenously added DNA.Abstract:
We have developed a cell-free system for studying the synthesis of mRNA in mammalian cells. The system consists of a dialyzed and concentrated whole-cell extract derived from HeLa cells, small molecules and cofactors needed for transcription, and exogenously added DNA. Accurate transcription by RNA polymerase II is entirely dependent upon addition of promoter-containing eukaryotic DNA. At optimal DNA and extract concentrations, transcription initiation from the adenovirus serotype 2 late promoter is readily detectable, and specific transcripts over 4000 nucleotides in length are observed. The RNA synthesized in vitro contains the same 5' capped RNase T1 undecanucleotide as does the in vivo transcript. RNA synthesis also initiates accurately at both an early and an intermediate adenovirus promoter site.read more
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Recombinant genomes which express chloramphenicol acetyltransferase in mammalian cells.
TL;DR: A series of recombinant genomes which directed expression of the enzyme chloramphenicol acetyltransferase (CAT) in mammalian cells provided a uniquely convenient system for monitoring the expression of foreign DNAs in tissue culture cells.
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Efficient in vitro synthesis of biologically active RNA and RNA hybridization probes from plasmids containing a bacteriophage SP6 promoter
TL;DR: In this paper, a simple and efficient method for synthesizing pure single stranded RNAs of virtually any structure is described, based on the unusually specific RNA synthesis by bacteriophage SP6 RNA polymerase which initiates transcription exclusively at an SP6 promoter.
Journal ArticleDOI
DNA sequence and expression of the B95-8 Epstein—Barr virus genome
R. Baer,R. Baer,Alan T. Bankier,Mark D. Biggin,Prescott L. Deininger,Paul J. Farrell,Toby J. Gibson,Graham F. Hatfull,Graham S. Hudson,Sandra C. Satchwell,C. Séguin,C. Séguin,P. S. Tuffnell,Bart Barrell +13 more
TL;DR: The complete (172,282 base pairs) nucleotide sequence of the B95-8 strain of Epstein–Barr virus has been established using the dideoxynucleotide/M13 sequencing procedure.
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The Rous sarcoma virus long terminal repeat is a strong promoter when introduced into a variety of eukaryotic cells by DNA-mediated transfection
TL;DR: The results indicate that the Rous sarcoma virus LTR can direct synthesis of high levels of functional mRNA and has a wide expression range.
Journal ArticleDOI
Mechanism of activation of a human oncogene.
Clifford J. Tabin,Scott M. Bradley,Cornelia I. Bargmann,Robert A. Weinberg,Alex G. Papageorge,Edward M. Scolnick,Ravi Dhar,Douglas R. Lowy,Esther H. Chang,Esther H. Chang +9 more
TL;DR: Experiments are presented that localize the genetic lesion that led to activation of the oncogene that affects the structure of theOncogene-encoded protein.
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