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互补的DNA编码点击甲虫光酶可以引起不同颜色的生物发光
K V Wood1, Y A Lam, H H Seliger
1Department of Chemistry, University of California, San Diego, La Jolla 92093.
Science (New York, N.Y.)
|May 12, 1989
概括
研究人员分离了点击甲虫互补DNA (cDNA) 克隆,这些克隆在细菌中产生可见生物发光. 这些克隆对四种不同的光酶进行编码,在分子生物学实验中作为多个记者基因有用.
科学领域:
- 生物化学 生物化学
- 分子生物学分子生物学
- 遗传学 是一个遗传学.
背景情况:
- 生物发光是一种在各种生物体中发现的自然发光过程.
- 露西法酶是催化生物发光反应的酶.
- 点击甲虫 (Pyrophorus plagiophthalamus) 拥有独特的生物发光特性.
研究的目的:
- 隔离和表征来自点击甲虫Pyrophorus plagiophthalamus的编码 luciferases的互补DNA (cDNA) 克隆.
- 评估这些克隆基因在分子生物学中作为记者基因的潜力.
主要方法:
- 使者RNA (mRNA) 从Pyrophorus plagiophthalamus的光器官中分离出来.
- 建立了互补DNA (cDNA) 库,并对11个克隆进行了测序.
- 分离的cDNA克隆被表达在大肠杆菌中,以评估它们的生物发光能力.
主要成果:
- 成功生成了11个cDNA克隆,它们能够在大肠杆菌表达时引起可见生物发光.
- 这些克隆编码了四种不同类型的光酶,产生绿色,黄色绿色,黄色和色光 (546-593nm).
- 这些甲虫 luciferases 分享 95-99% 的氨基酸序列相同性彼此,但只有 48% 与火虫 luciferase.
结论:
- 描述的点击甲虫光酶代表了生物发光应用的新工具.
- 这些光酶发出的鲜明的颜色使它们适合在记者基因分析中进行多重复合.
- 这些发现扩大了研究人员在各种生物研究中使用生物发光记者的工具包.
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