Sumikawa K, Houghton M, Smith J C, Bell L, Richards B M, Barnard E A
Nucleic Acids Res. 1982 Oct 11;10(19):5809-22. doi: 10.1093/nar/10.19.5809.
A rare cDNA coding for most of the alpha subunit of the Torpedo nicotinic acetylcholine receptor has been cloned into bacteria. The use of a mismatched oligonucleotide primer of reverse transcriptase facilitated the design of an efficient, specific probe for recombinant bacteria. DNA sequence analysis has enabled the elucidation of a large part of the polypeptide primary sequence which is discussed in relation to its acetylcholine binding activity and the location of receptor within the plasma membrane. When used as a radioactive probe, the cloned cDNA binds specifically to a single Torpedo mRNA species of about 2350 nucleotides in length but fails to show significant cross-hybridisation with alpha subunit mRNA extracted from cat muscle.
编码电鳐烟碱型乙酰胆碱受体大部分α亚基的一种罕见互补DNA已被克隆到细菌中。使用逆转录酶的错配寡核苷酸引物有助于设计一种用于重组细菌的高效、特异性探针。DNA序列分析已能够阐明多肽一级序列的大部分内容,并结合其乙酰胆碱结合活性以及受体在质膜中的位置进行了讨论。当用作放射性探针时,克隆的互补DNA特异性结合到一种长度约为2350个核苷酸的单一电鳐信使核糖核酸物种上,但与从猫肌肉中提取的α亚基信使核糖核酸未显示出明显的交叉杂交。