원문정보
Effects of Environmental Conditions on the Expression of Hantaan Viral Nucleocapsid Gene in Escherichia coli
초록
영어
Viruses belonging to the Hantavirus genus cause two acute severe illness in humans, i.e., Haemorrhagic Fever with Renal Syndrome (HFRS) and Hantavirus Pulmonary Syndrome(HPS). Among them, Hantaan virus is one of the most important viruses causing HFRS. Recombinant expression vectors, pKK-NP and pET-NP, with Hantaan viral nucleocapsid gene were constructed, and used to transform Eschericia coli BL21(DE3). Stability of the vectors in the host strain, and effects of some environmental conditions on the expression of nucleocapsid gene were studied. Expression vector, pKK-NP, was very unstable, and the expression level of nucleocapsid gene was very low compared to that of pET-NP. BL21(pET-NP) produced about 100 mg of N protein per liter of culture broth. Induction time did not show any significant difference on the expression level of nucleocapsid gen and cell growth. BL21(pET-NP) culture at 35℃ showed a little higher expression level than at 30℃ during growth phase, but reached to the same level at stationary phase. Total expression level was proportional to supplemented glucose concentration of media up to 0.5% along with cell growth, but expression level per unit cell mass was inversely proportional to glucose concentration and maximal when glucose was not supplemented at all.
목차
재료 및 방법
균주, 시약 및 재료
균체의 배양 및 파쇄
EIA에 의한 뉴클레오캡시드 단백질의 정량
플라스미드의 안정성 연구
결과 및 고찰
발현된 뉴클레오캡시드 단백질의 정량
플라스미드의 안정성 연구
유전자 재조합 균주의 생장 특성
온도에 따른 영향
Induction time의 영향
Glucose 농도에 따른 영향
요약
참고문헌