CAJ | 학술논문

用PVS -CP基因的原核表达载体pBAD -PVS转化受体菌 TOP10获得了重组菌 TOP10（pBAD -PVS），经阿拉伯糖诱导该重组菌表达出了46kDa的特异性融合蛋白，在Western blotting图谱上该蛋白与PVS特异性抗体（IgG）反应，呈现一条阳性杂交带，表明46kDa蛋白带是PVS -CP基因正确表达的产物。从重组菌中提取包涵体，包涵体经过4mol/L尿素洗涤后用超纯水4℃过夜溶解，所获溶液即为高纯度 PVS重组CP水溶液，也就是说本研究建立了一种获得高纯度重组CP的非常简便的方法。
용PVS -CP기인적원핵표체재체pBAD -PVS전화수체균 TOP10획득료중조균 TOP10（pBAD -PVS），경아랍백당유도해중조균표체출료46kDa적특이성융합단백，재Western blotting도보상해단백여PVS특이성항체（IgG）반응，정현일조양성잡교대，표명46kDa단백대시PVS -CP기인정학표체적산물。종중조균중제취포함체，포함체경과4mol/L뇨소세조후용초순수4℃과야용해，소획용액즉위고순도 PVS중조CP수용액，야취시설본연구건립료일충획득고순도중조CP적비상간편적방법。
The prokaryotic expression vector of CP gene of potato virus S (pBAD -PVS) was introduced into E .coli strain TOP10 ,then the recombinant strain TOP10 (pBAD-PVS) was induced with L -arabi-nose ,and a 46kDa specific protein band was found in SDS pattern of total protein .In Western blotting a-nalysis ,the 46kDa protein reacted with PVS -IgG ,and a positive band was observed ,the result indicated that the 46kDa protein was the correct expression product of PVS -CP gene .Inclusion body was extracted from TOP10 (pBAD -PVS) and washed with 4 mol/L urea .The washed inclusion body was suspended in ultra pure water and kept at 4℃ overnight .The resulting solution was indeed the high -purity recombi-nant CP ,a very simple purification method of PVS recombinant CP was established .