4 讨论
洛伐他汀是目前治疗高血脂症的常用药物之一。国内工业化生产洛伐他汀的效率并不理想,以致研究者们纷纷以增产洛伐他汀为目标开展多方面的研究。目前从基因工程方面着手改善其产量成为热点。本文克隆的lovE和mkH基因编码产物是一个洛伐他汀生物合成的调控蛋白或转录因子,对其深入研究将有助于理解和控制洛伐他汀生物合成。
Kennedy、Park等将lovE基因突变失活导致检测不到任何洛伐他汀中间产物,同时将一个额外拷贝的lovE转化土曲霉,洛伐他汀产量增加5~7倍[10,16]。从上述数据可以看出对lovE和mkH基因以及其编码蛋白的结构功能的进一步研究将为深入理解洛伐他汀生物合成及其遗传调控方面提供参考,且为进一步表达基因、纯化相关蛋白、制备多克隆抗体、研究基因体外转录、转录因子与DNA的体外模拟结合以深入了解该类转录因子生物学功能和作用机理奠定了实验基础。
本研究利用国际互联网生物信息学资源,通过计算分析两蛋白物理化学特性,得知此两核蛋白和一些转录因子以及调控序列编码产物有共同的GAL4类锌指结构保守序列区。相似性搜索比对得知lovE和mkH不论基因还是编码产物序列都具有高度相似性,存在共同的DNA结合保守序列。
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