We have developed a procedure for preparing extracts from nuclei of human tissue culture cells that directs accurate transcription initiation in vitro from class II promoters. Conditions of extraction and assay have been optimized for maximum activity using the major late promoter of adenovirus 2. The extract also directs accurate transcription initiation from other adenovirus promoters and cellular promoters. The extract also directs accurate transcription initiation from class III promoters (tRNA and Ad 2 VA).

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Accurate transcription initiation by RNA polymerase II in a soluble extract from isolated mammalian nuclei

Detection of two restriction endonuclease activities in Haemophilus parainfluenzae using analytical agarose--ethidium bromide electrophoresis

A widely recognized difficulty of presently used methods for cDNA cloning is obtaining cDNA segments that contain the entire nucleotide sequence of the corresponding mRNA. The cloning procedure described here mitigates this shortcoming. Of the 10(5) plasmid-cDNA recombinants obtained per microgram of rabbit reticulocyte mRNA, about 10% contained a complete alpha- of beta-globin mRNA sequence, and at least 30 to 50%, but very likely more, contained the entire globin coding regions. We attribute the high efficiency of cloning full- or nearly full-length cDNA to (i) the fact that the plasmid DNA vector itself serves as the primer for first- and second-strand cDNA synthesis, (ii) the lack of any nuclease treatment of the products, and (iii) the fact that one of the steps in the procedure results in preferential cloning of recombinants with full-length cDNA's over those with truncated cDNA's.

High-efficiency cloning of full-length cDNA.

https://mmbr.asm.org/content/mmbr/47/1/1.full.pdf

Comparison of initiation of protein synthesis in procaryotes, eucaryotes, and organelles.

Splicing of Messenger RNA Precursors

Transcription of Simian virus 40. I : separation of the strands of SV40 DNA and hybridization of the separated strands to RNA extracted from lytically infected and transformed cells

https://www.cell.com/cell/pdf/0092-8674(79)90100-4.pdf

Nucleotide sequence analysis of the leader segments in a cloned copy of adenovirus 2 fiber mRNA

Mammalian deoxyribonucleic acid-dependent ribonucleic acid polymerases. I. Purification and properties of an -amanitin-sensitive ribonucleic acid polymerase and stimulatory factors from HeLa and KB cells.

Walter Keller

Papers

Transcription of Simian virus 40. I : separation of the strands of SV40 DNA and hybridization of the separated strands to RNA extracted from lytically infected and transformed cells

Nucleotide sequence analysis of the leader segments in a cloned copy of adenovirus 2 fiber mRNA

Mammalian deoxyribonucleic acid-dependent ribonucleic acid polymerases. I. Purification and properties of an -amanitin-sensitive ribonucleic acid polymerase and stimulatory factors from HeLa and KB cells.

Cleavage Maps of DNA from Adenovirus Types 2 and 5 by Restriction Endonucleases EcoRI and HpaI

Extent of Terminal Repetition in Adenovirus 2 DNA