Cloning of Acetylcholinesterase Gene in a Microbial Vector
Abstract
Acetylcholinesterase is the major site of action for several of the nerve gasses. It is therefore essential to elucidate the structure of the acetylcholinesterase molecule, in order to devise ways to protect against these poisons. This report describes attempts to obtain a cDNA clone encoding the acetylcholinesterase molecule. Such a clone would be useful because the primary amino acid sequence of the enzyme could be inferred from the cloned DNA sequence, and it could ultimately be used to produce sufficient quantities of the enzyme to determine its three dimensional structure. The electric organ from Torpedo californica, which produces high levels of acetycholinesterase, was used as the mRNA source for the recombinant DNA work. Identification of the mRNA encoding acetylcholinesterase (approximately .5% of the total mRNA) was accomplished by immuno-precipitation of radio-labeled in vitro translation products using an antiserum specific for the enzyme. The mRNA for the enzyme was characterized and enriched by fractionation on sucrose gradients. cDNA molecules were synthesized using both enriched mRNA and total RNA, and these were cloned in the plasmid vector, pBR322. (js)
Document Details
- Document Type
- Technical Report
- Publication Date
- Oct 19, 1984
- Accession Number
- ADA226191
Entities
People
- Brian L. West
- John D. Baxter
Organizations
- University of California, San Francisco