CDNA cloning and characterization of a secreted luciferase from the luminous Japanese ostracod, Cypridina noctiluca
CDNA cloning and characterization of a secreted luciferase from the luminous Japanese ostracod, Cypridina noctiluca
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DOI：
10.1271/bbb.68.565
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发表时间：
2004-03-01
影响因子：
1.6
通讯作者：
Ohmiya, Y
中科院分区：
文献类型：
--
作者：
Nakajima, Y;Kobayashi, K;Ohmiya, Y
海洋介形类 Vargula hilgendorfii 分泌的荧光素酶是活体哺乳动物细胞基因表达测定的有用工具。我们从栖息于日本海岸的介形类Cypridina noctiluca 中克隆了一种新的分泌型荧光素酶的cDNA。 C. noctiluca 荧光素酶由 553 个氨基酸残基组成，根据核苷酸序列推断，分子量为 61,415 Da。与希尔根多夫弧菌荧光素酶的核苷酸和氨基酸序列同源性分别为79.2%和83.1%。夜光藻荧光素酶可以在培养的哺乳动物细胞中表达并分泌。表达的 C noctiluca 荧光素酶的特性与 V. hilgendorfii 荧光素酶的特性相似。然而，培养基中夜光藻荧光素酶的活性远高于希尔根多夫弧菌荧光素酶，表明夜光藻荧光素酶是一种高效的报告酶，可用于实时、连续监测活细胞中的基因表达。
A secreted luciferase from the marine ostracod, Vargula hilgendorfii, is a useful tool for gene expression assays in living mammalian cells. We have cloned the cDNA of a new secreted luciferase from the ostracod Cypridina noctiluca, which inhabits the coast of Japan. C. noctiluca luciferase consists of 553 amino acid residues with a molecular mass of 61,415 Da, as deduced from the nucleotide sequence. The homologies of nucleotide and amino acid sequences with V. hilgendorfii luciferase are 79.2% and 83.1%, respectively. C. noctiluca luciferase can expressed in and secreted from cultured mammalian cells. The characteristic properties of expressed C noctiluca luciferase are similar to those of V. hilgendorfii luciferase. However, the activity of C. noctiluca luciferase in culture medium is much higher than that of V. hilgendorfii luciferase, suggesting that C noctiluca luciferase is a highly potent reporter enzyme for real-time and continuous monitoring of gene expression in living cells.