Lorenz W W, McCann R O, Longiaru M, Cormier M J
Department of Biochemistry, University of Georgia, Athens 30602.
Proc Natl Acad Sci U S A. 1991 May 15;88(10):4438-42. doi: 10.1073/pnas.88.10.4438.
Renilla reniformis is an anthozoan coelenterate capable of exhibiting bioluminescence. Bioluminescence in Renilla results from the oxidation of coelenterate luciferin (coelenterazine) by luciferase [Renilla-luciferin:oxygen 2-oxidoreductase (decarboxylating), EC 1.13.12.5]. In vivo, the excited state luciferin-luciferase complex undergoes the process of nonradiative energy transfer to an accessory protein, green fluorescent protein, which results in green bioluminescence. In vitro, Renilla luciferase emits blue light in the absence of any green fluorescent protein. A Renilla cDNA library has been constructed in lambda gt11 and screened by plaque hybridization with two oligonucleotide probes. We report here the isolation and characterization of a luciferase cDNA and its gene product. The recombinant luciferase expressed in Escherichia coli is identical to native luciferase as determined by SDS/PAGE, immunoblot analysis, and bioluminescence emission characteristics.
海肾（Renilla reniformis）是一种能够产生生物发光的珊瑚虫纲腔肠动物。海肾中的生物发光是由荧光素酶[海肾荧光素：氧2-氧化还原酶（脱羧），EC 1.13.12.5]氧化腔肠动物荧光素（腔肠素）所致。在体内，激发态的荧光素-荧光素酶复合物经历非辐射能量转移过程至一种辅助蛋白——绿色荧光蛋白，从而产生绿色生物发光。在体外，海肾荧光素酶在没有任何绿色荧光蛋白的情况下发出蓝光。已构建了一个λgt11海肾cDNA文库，并用两个寡核苷酸探针通过噬菌斑杂交进行筛选。我们在此报告荧光素酶cDNA及其基因产物的分离和特性。通过SDS/PAGE、免疫印迹分析和生物发光发射特性测定，在大肠杆菌中表达的重组荧光素酶与天然荧光素酶相同。
