Funahashi Aki, Itakura Takao, Hassanin Abeer A I, Komatsu Masaharu, Hayashi Seiichi, Kaminishi Yoshio
Faculty of Fisheries, Kagoshima University, 4-50-20 Shimoarata, Kagoshima 890-0056, Japan.
J Genet. 2017 Mar;96(1):127-133. doi: 10.1007/s12041-017-0751-5.
In this study, the localization of fluorescent protein (FP) was characterized in the muscles of four species and two subspecies of eels Anguilla anguilla, A. australis, A. bicolor bicolor (b.), A. bicolor pacifica (p.) and A. mossambica in addition to the previously reported A. japonica. The open reading frame of each eel FP was 417 bp encoding 139 amino acid residues. The deduced amino acid sequences among the four species and two subspecies exhibited 91.4-100% identity, and belonged to the fatty-acid-binding protein (FABP) family. The gene structure of eel FPs in A. japonica, A. anguilla, A. australis, A. bicolor b., A. bicolor p. and A. mossambica have four exons and three introns, and were common to that of FABP family. The apo eel FPs expressed by Escherichia coli with recombinant eel FP genes were analysed for the fluorescent properties in the presence of bilirubin. The excitation and emission spectra of holo eel FPs had the maximum wavelengths of 490-496 and 527-530 nm, respectively. The holo eel FPs indicated that the fluorescent intensities were stronger in A. japonica and A. bicolor than in A. mossambica, A. australis and A. anguilla. The comparison of amino acid sequences revealed two common substitutions in A. mossambica, A. australis and A. anguilla with weak fluorescent intensity.
在本研究中,除了先前报道的日本鳗鲡外,还对欧洲鳗鲡、澳大利亚鳗鲡、双色鳗鲡指名亚种、双色鳗鲡太平洋亚种和莫桑比克鳗鲡这四个物种和两个亚种的肌肉中荧光蛋白(FP)的定位进行了表征。每种鳗鲡FP的开放阅读框为417 bp,编码139个氨基酸残基。这四个物种和两个亚种之间推导的氨基酸序列显示出91.4 - 100%的同一性,并且属于脂肪酸结合蛋白(FABP)家族。日本鳗鲡、欧洲鳗鲡、澳大利亚鳗鲡、双色鳗鲡指名亚种、双色鳗鲡太平洋亚种和莫桑比克鳗鲡中鳗鲡FP的基因结构有四个外显子和三个内含子,这与FABP家族的基因结构相同。用重组鳗鲡FP基因在大肠杆菌中表达的脱辅基鳗鲡FP在胆红素存在的情况下进行了荧光特性分析。全鳗鲡FP的激发光谱和发射光谱的最大波长分别为490 - 496 nm和527 - 530 nm。全鳗鲡FP表明,日本鳗鲡和双色鳗鲡中的荧光强度比莫桑比克鳗鲡、澳大利亚鳗鲡和欧洲鳗鲡中的更强。氨基酸序列比较揭示了荧光强度较弱的莫桑比克鳗鲡、澳大利亚鳗鲡和欧洲鳗鲡中有两个共同的替换。