20090319 黑曲霉基因组及柠檬酸生产菌种改造.pptVIP

高温发酵菌株的高通量筛选 全自动化：平皿制备、涂布、平板初筛、点种、液体接种、发酵、样品处理、分析 高效率：几十万克隆每天 黑曲霉基因组简介和柠檬酸菌种改造初步方案 孙际宾李寅中国科学院工业生物技术研究所 (筹) -------- 谢谢！--------- * * * 略提一下，后面讲 * 比较基因组学获得基因组成（增减、沉默）、调控区、蛋白质序列(修饰位点）的改变 比较转录组学获得两种菌株在相同环境条件下基因表达方面质和量的差异 与代谢网络分析结合，获得网络调控的动态：在高产条件下，哪些途径该加强，哪些该减弱，道理何在，其调控因子是什么？ 反向工程分析time series基因表达数据，获得调控网络 哪些途径可以进一步改造（如切除分支途径） 比较蛋白组研究 蛋白质组在各细胞分区的动态变化 运输系统是否发生变化？ 哪些蛋白质、怎样被修饰？是否与目的产物产生酶水平调控相关？ 比较代谢组和代谢流组 哪些途径存在瓶颈 哪些代谢途径流向副产物形成 … … * * * * * This stimulation of citrate accumulation by oxygen is paralleled by the induction of the cyanide-resistant, salicylhydroxamic acid-sensitive alternative oxidase, which allows the sites of energy conservation to be bypassed and decreases the energy yield of the cells (Zehentgruber et al. 1980; Kirimura et al. 2000). \ The physiological function of this uncoupled respiration is thought to be the removal of the reducing equivalents in excess. The almost quantitative conversion of hexose to citric acid results in a net production of one mole of ATP and three moles of NADH, whose turn-over is limited by the absence of biosynthetic processes of equivalent capacity. Hence, the cells do not require much ATP and a switch from cytochrome-dependent respiration to the alternative route would enable the fungus to reoxidize its NADH without concomitant ATP production. The alternative respiratory pathway has also been suggested to contribute to the prevention of the generation of reactive oxygen species, which are formed when the cytochrome path is impaired and/or the intracellular oxygen is in excess (Popov et al. 1997; Maxwell et al. 1999; Karaffa et al. 2001) and can be considered as an integral part of the antioxidant defense system in eukaryotes. * Biotechnology is going out of the dark era. * * 生物信息学优势 * 黑曲霉基因组简介和柠檬酸菌种改造初步方案 孙际宾李寅中国科学院工业生物技术研究所 (筹) 实例：基因组重测指导赖氨酸高产菌株构建 Ohnishi et al. 2002 比较基因组学 3个点突变?野生型菌株： 0?80g/l 50h?27h 1.解密黑曲霉基因组 Nick Read 1991 黑曲霉测序大事记 2000.07. 荷兰帝斯曼DSM公司启动了对其拥有的黑曲霉菌株CBS 513.88的测序计划，采用BAC加shortgun法 2001.12