【摘要】：目的 觀察非分型流感嗜血桿菌ATCC49247的生長條件中的因子需求情況，優(yōu)化非分型流感嗜血桿Haps蛋白質(zhì)片段表達與分泌條件，并用弱陽離子交換層析柱純化Hap_s蛋白質(zhì)片段，優(yōu)化Hap_s蛋白質(zhì)片段的洗脫條件，包括pH值和離子強度，為進一步研究打下基礎(chǔ)。 方法 1．將非分型流感嗜血桿菌ATCC49247均勻涂布于LB平板培養(yǎng)基上，并在涂菌后的LB平板培養(yǎng)基上放置三張無菌濾紙，在濾紙上分別滴入20μl的氯化血紅素液體、β-NAD液體及含有氯化血紅素和β-NAD的混合液體，然后將平板置37℃孵箱孵育。 2．用心腦浸液液體培養(yǎng)基(sBHI)培養(yǎng)非分型流感嗜血桿菌ATCC49247，觀察在不同培養(yǎng)時間段，非分型流感嗜血桿菌培養(yǎng)液在600nm波長的光吸收值(A_(600))變化，并用SDS-PAGE電泳檢測Hap_s蛋白質(zhì)片段分泌量的變化。 3．非分型流感嗜血桿菌ATCC49247在心腦浸液液體培養(yǎng)基(sBHI)中37℃培養(yǎng)18小時后，，加入IPTG誘導不同時間，觀察其對Haps蛋白質(zhì)片段分泌量的影響。
[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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