Functional properties of strychnine-sensitive glycine receptors expressed in Xenopus oocytes injected with a single mRNA

Abstract

Mature rat spinal cord cDNA libraries constructed in lambda gt10 and lambda ZAPII were screened with an oligonucleotide probe (39 mer), and 4 clones that possess DNA-inserts encoding a glycine receptor subunit were obtained. The cloned cDNAs were used to reconstruct the nucleotide sequence of the full-length open reading frame consisting of 1350 base pairs (bp) as well as the 5'-(184 bp) and 3'-(591 bp) non-coding regions. Synthetic RNA transcribed in vitro from the glycine receptor cDNA induced Xenopus oocytes to synthesize functional glycine receptor that generated large Cl- currents. The electrophysiological properties of the wild-type receptor and some mutant receptors produced by site-directed mutagenesis were analyzed.