非分型流感嗜血杆菌培养条件的优化及Hap_s蛋白的分离纯化

发布时间：2019-03-14 13:31

【摘要】：目的 观察非分型流感嗜血杆菌ATCC49247的生长条件中的因子需求情况，优化非分型流感嗜血杆Haps蛋白质片段表达与分泌条件，并用弱阳离子交换层析柱纯化Hap_s蛋白质片段，优化Hap_s蛋白质片段的洗脱条件，包括pH值和离子强度，为进一步研究打下基础。 方法 1．将非分型流感嗜血杆菌ATCC49247均匀涂布于LB平板培养基上，并在涂菌后的LB平板培养基上放置三张无菌滤纸，在滤纸上分别滴入20μl的氯化血红素液体、β-NAD液体及含有氯化血红素和β-NAD的混合液体，然后将平板置37℃孵箱孵育。 2．用心脑浸液液体培养基(sBHI)培养非分型流感嗜血杆菌ATCC49247，观察在不同培养时间段，非分型流感嗜血杆菌培养液在600nm波长的光吸收值(A_(600))变化，并用SDS-PAGE电泳检测Hap_s蛋白质片段分泌量的变化。 3．非分型流感嗜血杆菌ATCC49247在心脑浸液液体培养基(sBHI)中37℃培养18小时后，，加入IPTG诱导不同时间，观察其对Haps蛋白质片段分泌量的影响。
[Abstract]:Objective to observe the demand for factors in the growth conditions of Haemophilus influenzae ATCC49247, and to optimize the expression and secretion conditions of Haps protein fragments in non-typing Haemophilus influenzae. The Hap_s protein fragment was purified by weak cation exchange chromatography, and the elution conditions of Hap_s protein fragment, including pH value and ionic strength, were optimized to lay a foundation for further study. The non-typing Haemophilus influenzae ATCC49247 was uniformly coated on the LB plate medium and three sterile filter papers were placed on the LB plate medium after the strain was coated. A 20 渭 l hemin liquid, a 尾-NAD liquid and a mixed liquid containing hemin and 尾-NAD were added to the filter paper respectively. Then the plates were incubated in a 37 鈩

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