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用于3',5'-环磷酸腺苷及环核苷酸磷酸二酯酶抑制剂的琼脂平板筛选程序

Agar plate screening procedure for cyclic adenosine 3',5'-monophosphate and inhibitors of cyclic nucleotide phosphodiesterase.

作者信息

Somers P J, Higgens C E

出版信息

Appl Environ Microbiol. 1977 Nov;34(5):604-6. doi: 10.1128/aem.34.5.604-606.1977.

DOI:10.1128/aem.34.5.604-606.1977
PMID:201216
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC242711/
Abstract

A procedure is described for the semiquantitative measurement of cyclic adenosine 3',5'-monophosphate (cAMP) and detection of inhibitors of cAMP phosphodiesterase by an agar plate test. The assay organism was an adenyl cyclase-deficient mutant derived from Escherichia coli HfrH. In the presence of an acid base indicator, acid production from barbohydrate metabolism was observed as a yellow zone around filter paper disks containing cAMP. Since yellow zone formation reflects the presence of cAMP, a phosphodiesterase inhibitor can be detected indirectly by the presence of a yellow zone on assay plates from a reaction mixture of an inhibitor, phosphodiesterase, and cAMP. Three known cyclic nucleotide phosphodiesterase inhibitors were active against beef brain phosphodiesterase in this system.

摘要

本文描述了一种通过琼脂平板试验对环磷腺苷（cAMP）进行半定量测量以及检测cAMP磷酸二酯酶抑制剂的方法。测定用生物体是源自大肠杆菌HfrH的腺苷酸环化酶缺陷型突变体。在酸碱指示剂存在的情况下，碳水化合物代谢产生的酸在含有cAMP的滤纸圆盘周围呈现为黄色区域。由于黄色区域的形成反映了cAMP的存在，因此磷酸二酯酶抑制剂可通过抑制剂、磷酸二酯酶和cAMP反应混合物在测定平板上出现黄色区域而间接检测到。在该系统中，三种已知的环核苷酸磷酸二酯酶抑制剂对牛脑磷酸二酯酶具有活性。

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本文引用的文献

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New inhibitor of phosphodiesterase with anti-bronchoconstrictor properties.具有抗支气管收缩特性的新型磷酸二酯酶抑制剂。
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Investigation of the mechanism of action of the lipolytic agent 4-(3,4-dimethoxybenzyl)-2-imidazolidinone (Ro 7-2956).
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Requirement of adenosine 3', 5'-cyclic phosphate for flagella formation in Escherichia coli and Salmonella typhimurium.大肠杆菌和鼠伤寒沙门氏菌中鞭毛形成对3'，5'-环磷酸腺苷的需求。
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Regulation of lac messenger ribonucleic acid synthesis by cyclic adenosine 3',5'-monophosphate and glucose.3',5'-环磷酸腺苷和葡萄糖对乳糖信使核糖核酸合成的调控
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Cyclic 3',5'-nucleotide phosphodiesterase. Preparation of a partially inactive enzyme and its subsequent stimulation by snake venom.环3',5'-核苷酸磷酸二酯酶。部分失活酶的制备及其随后被蛇毒激活的过程。
Biochim Biophys Acta. 1969 Nov 4;191(2):303-15. doi: 10.1016/0005-2744(69)90249-6.
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Pleiotropic deficiency of carbohydrate utilization in an adenyl cyclase deficient mutant of Escherichia coli.大肠杆菌腺苷酸环化酶缺陷型突变体中碳水化合物利用的多效性缺陷
Biochem Biophys Res Commun. 1969 Sep 24;37(1):151-7. doi: 10.1016/0006-291x(69)90893-6.
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The stimulatory effect of cyclic adenosine 3'5'-monophosphate on DNA-directed synthesis of beta-galactosidase in a cell-free system.环磷酸腺苷在无细胞体系中对β-半乳糖苷酶的DNA指导合成的刺激作用。
Proc Natl Acad Sci U S A. 1969 May;63(1):118-22. doi: 10.1073/pnas.63.1.118.
10
The role of the lac promotor locus in the regulation of beta-galactosidase synthesis by cyclic 3',5'-adenosine monophosphate.乳糖启动子位点在3',5'-环磷酸腺苷对β-半乳糖苷酶合成的调控中的作用。
Proc Natl Acad Sci U S A. 1968 Dec;61(4):1336-42. doi: 10.1073/pnas.61.4.1336.