Luc genes: introduction of colour into bioluminescence assays.-猫眼课题宝

Luc 基因：将颜色引入生物发光测定。

DOI：

发表时间：

1990

期刊：

Journal of bioluminescence and chemiluminescence (Print)

影响因子：

通讯作者：

中科院分区：

文献类型：

作者：

Keith V. Wood

发光测定通常仅基于光强度的测量。将颜色作为测定的附加参数包括在内可以增加信息内容。发光的颜色变化在甲虫发光酶中特别普遍。为了研究酶的结构和颜色之间的关系，从Jamalcan点击甲虫的酶作为模型系统进行了检查。这些发光酶的发光范围从绿色到橙子，尽管它们的氨基酸序列差异小于5%。通过突变它们各自的cDNA克隆，鉴定了负责颜色变化的氨基酸。这些特定的氨基酸很少，它们对颜色的作用与酶的结构无关。对它们的影响的分析表明，甲虫荧光素酶之间颜色变化的潜力大于点击甲虫荧光素酶之间的明显差异。由于影响颜色的酶结构的微妙变化，发射不同颜色的酶可能用作成对的遗传报告基因。它们应与宿主的细胞内环境等效地相互作用，但在其测定中可通过颜色区分。这种配对的报告基因可用于观察同时发生的事件，或为发光测量提供内部对照。

Luminescence assays are generally based on measurements of light intensity alone. Inclusion of colour as an additional parameter of the assay could increase the information content. Colour variation in luminescence is particularly prevalent among beetle luciferases. To study the relationship between enzyme structure and colour, luciferases from a Jamalcan click beetle were examined as a model system. These luciferases emit light ranging from green to orange, though their amino acid sequences differ by less than 5%. Through mutation of their respective cDNA clones, the amino acids responsible for the colour variation were identified. These specific amino acids are few, and they act upon colour independently with respect to the enzyme structure. Analysis of their effects indicates that the potential for colour variation among beetle luciferases is greater than is evident among the click beetle luciferase. Because of the subtle changes of enzyme structure that effect colour, luciferases that emit different colours may be useful as paired genetic reporters. They should interact equivalently with the intracellular environment of a host, but could be distinguished by colour in their assay. Such paired reporters could be used to observed simultaneous events, or to provide internal control for luminescence measurements.