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首页> 外文会议>Progress on post-genome technologies and modern natural products >MOLECULAR BREEDING OF LUCIFERASE SUITABLE FOR PYROSEQUENCING BY CELL SURFACE ENGINEERING
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MOLECULAR BREEDING OF LUCIFERASE SUITABLE FOR PYROSEQUENCING BY CELL SURFACE ENGINEERING

机译：适用于细胞表面工程热测序的核糖核酸酶分子育种

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There are increasing needs of higher-speed and lowercost Next Generation DNA sequencing system. One of the Next Generation DNA sequencing systems is a pyrosequence system, which use luciferase. However, there is a crucial problem facing current state of pyrosequencing system. The pyrosequencing system is forced to adopt dATP analog, because of ambiguous substrate recognition of lucif erase, and dATP analog is expensive. If lucif erase can recognize the difference between ATP and dATP, the pyrosequencing system will avoid adopting dATP analog. To improve these problems, we tried to create a novel lucif erase with high ATP-specificity. At first, we selected two amino acids from whole lucif erase sequences and randomized these two amino acids. Mutant luciferase library was constructed by yeast cell surface engineering. Through two screening steps, we isolated candidate mutants and analyzed these mutants. Consequently, we succeeded in obtaining luciferase with increased ATP-specificity and -reactivity.
机译：对高速和低成本下一代DNA测序系统的需求不断增长。下一代DNA测序系统之一是使用荧光素酶的焦磷酸测序系统。然而，焦磷酸测序系统的当前状态面临着一个关键问题。由于荧光清除的底物模棱两可，焦磷酸测序系统被迫采用dATP类似物，而dATP类似物价格昂贵。如果lucif擦除可以识别ATP和dATP之间的差异，则焦磷酸测序系统将避免采用dATP类似物。为了改善这些问题，我们尝试创建一种具有高ATP特异性的新型lucif擦除。首先，我们从完整的lucif擦除序列中选择了两个氨基酸，并将这两个氨基酸随机化。通过酵母细胞表面工程构建突变体荧光素酶文库。通过两个筛选步骤，我们分离了候选突变体并分析了这些突变体。因此，我们成功获得了具有增加的ATP特异性和-反应性的荧光素酶。

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