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电鳐乙酰胆碱酯酶催化亚基cDNA克隆的分离

Isolation of a cDNA clone for a catalytic subunit of Torpedo marmorata acetylcholinesterase.

作者信息

Sikorav J L, Vallette F, Grassi J, Massoulié J

出版信息

FEBS Lett. 1985 Dec 2;193(2):159-63. doi: 10.1016/0014-5793(85)80142-3.

Abstract

We have constructed a cDNA library from Torpedo marmorata electric organ poly(A+) RNA in the lambda phage expression vector lambda gt11. This library has been screened with polyclonal anti-acetylcholinesterase antibodies. One clone, lambda AChE1, produced a fusion protein which was recognized by the antibodies and which prevented the binding of native acetylcholinesterase in an enzymatic immune assay. These results indicate that lambda AChE1 contains a cDNA insert coding for a part of a catalytic subunit of Torpedo acetylcholinesterase. The 200-base-pair cDNA insert hybridized to three mRNAs (14.5, 10.5 and 5.5 kb) from Torpedo electric organs. These mRNAs were also detected in Torpedo electric lobes.

摘要

我们用λ噬菌体表达载体λgt11构建了来自电鳐电器官多聚腺苷酸(poly(A+))RNA的cDNA文库。该文库已用多克隆抗乙酰胆碱酯酶抗体进行筛选。一个克隆λAChE1产生了一种融合蛋白,该蛋白能被抗体识别,并在酶免疫测定中阻止天然乙酰胆碱酯酶的结合。这些结果表明,λAChE1含有一个编码电鳐乙酰胆碱酯酶催化亚基一部分的cDNA插入片段。这个200个碱基对的cDNA插入片段与来自电鳐电器官的三种mRNA(14.5、10.5和5.5 kb)杂交。这些mRNA在电鳐电叶中也被检测到。

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