Establishing a Stable Expression System for Studies of Acetylcholine Receptors

Toni Claudio, Steven Sine, F. J. Sigworth, Henry L. Paulson, Deborah Hartman

Research output: Contribution to journalArticle

2 Scopus citations

Abstract

The chapter uses two methods for introducing the acetylcholine receptor (AChR) subunit cDNAs into cultured cells: DNA-mediated gene transfer using calcium phosphate precipitation and retroviral infection. These two methods were chosen because of the properties peculiar to, but not necessarily unique to, the AChR. One consideration was the need to stably introduce four different cDNAs into the same cell; another was the goal of introducing the cDNAs into muscle cell lines. Using DNA-mediated gene transfer, the chapter demonstrates that the cotransformation efficiency of introducing all four AChR cDNAs plus the selectable marker gene into the same Ltk-aprt-fibroblast cell was 80%. In addition, half of these cells took up approximately equal copy numbers of each cDNA. Thus, DNA-mediated gene transfer is a technique that will readily allow the introduction of multiple cDNAs into the genome of the same cell.

Original languageEnglish (US)
Pages (from-to)219-247
Number of pages29
JournalCurrent Topics in Membranes and Transport
Volume33
Issue numberC
DOIs
StatePublished - Jan 1988

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ASJC Scopus subject areas

  • Molecular Biology
  • Cell Biology

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