Alipour Bagher Said, Hosseinkhani Saman, Nikkhah Maryam, Naderi-Manesh Hossein, Chaichi Mohammad Javad, Osaloo Shahrokh Kazempour
Department of Biochemistry, Faculty of Basic Sciences, Tarbiat Modarres University, Tehran, Iran.
Biochem Biophys Res Commun. 2004 Dec 3;325(1):215-22. doi: 10.1016/j.bbrc.2004.10.022.
The first cDNA from lampyridae encoding a glow-worm luciferase from lantern mRNA of Lampyris turkestanicus has been cloned, sequenced, the amino acid sequence predicted, and the sequence reported to GenBank. The cDNA was 1644 base pairs in length and coding a 547-residue protein. The deduced amino acid sequence of the luciferase gene of L. turkestanicus showed 98.7% and 95.8% identity to Lampyris noctiluca and Pyrocoelia rufa, respectively. Phylogenetic analysis further confirmed that the deduced amino acid sequences of L. turkestanicus luciferase gene belong to the same subfamily, Lampyrinae. The cDNA encoding the luciferase of L. turkestanicus was expressed as a 62kDa band in recombinant Escherichia coli and showed green luminescence in the presence of luciferin. Amongst amino acid differences of L. turkestanicus and L. noctiluca (its clade) there are two important substitutions. Signature amino-acid sequences and motifs found in the deduced sequence are CK2-phospho site, ASN-glycosylation, myristoylation site, PKC-phospho site, microbodies C-terminal targeting signal, and AMP-binding domain.
首个来自光萤科的cDNA已被克隆、测序、预测氨基酸序列并提交至GenBank，该cDNA编码来自突厥光萤（Lampyris turkestanicus）灯笼mRNA的萤火虫荧光素酶。该cDNA长度为1644个碱基对，编码一个由547个残基组成的蛋白质。突厥光萤荧光素酶基因推导的氨基酸序列与夜光萤（Lampyris noctiluca）和红胸窗萤（Pyrocoelia rufa）分别具有98.7%和95.8%的同一性。系统发育分析进一步证实，突厥光萤荧光素酶基因推导的氨基酸序列属于同一亚科——光萤亚科。编码突厥光萤荧光素酶的cDNA在重组大肠杆菌中表达为一条62kDa的条带，并在荧光素存在的情况下发出绿色荧光。在突厥光萤和夜光萤（其进化枝）的氨基酸差异中有两个重要的替换。在推导序列中发现的特征性氨基酸序列和基序有CK2磷酸化位点、天冬酰胺糖基化位点、肉豆蔻酰化位点、蛋白激酶C磷酸化位点、微体C末端靶向信号和AMP结合结构域。
