CAJ | 학술논문

一段长度为880?bp的庚型肝炎病毒cDNA在大肠杆菌BL21(DE3)菌株中得到表达。此 cDNA被插入到表达质粒pGEX-5X-1中，位于编码日本血吸虫谷胱甘肽硫转移酶（GST）的DNA序列下游，并与GST处于同一阅读框。用乳糖在37℃下诱导表达出以包涵体形式存在的GST-NS53融合蛋白，并用脲溶法提取了该蛋白；在20℃诱导时，表达出的蛋白大部分可溶，用谷胱甘肽Sepharose-4B亲和层析柱对可溶性的融合蛋白进行了纯化。免疫印迹实验证明，此融合蛋白能被庚型肝炎病人的血清和自制的抗GST血清特异性地识别。用PC gene 软件对NS53氨基酸序列的亲水性和抗原决定簇进行了分析。本研究为庚型肝炎ELISA诊断试剂研制打下了基础。
일단장도위880?bp적경형간염병독cDNA재대장간균BL21(DE3)균주중득도표체。차 cDNA피삽입도표체질립pGEX-5X-1중，위우편마일본혈흡충곡광감태류전이매（GST）적DNA서렬하유，병여GST처우동일열독광。용유당재37℃하유도표체출이포함체형식존재적GST-NS53융합단백，병용뇨용법제취료해단백；재20℃유도시，표체출적단백대부분가용，용곡광감태Sepharose-4B친화층석주대가용성적융합단백진행료순화。면역인적실험증명，차융합단백능피경형간염병인적혈청화자제적항GST혈청특이성지식별。용PC gene 연건대NS53안기산서렬적친수성화항원결정족진행료분석。본연구위경형간염ELISA진단시제연제타하료기출。
A 880 bp cDNA localized to the putative NS5 region of HGV genome was expressed in E.coli BL21(DE3). The cDNA fragment was inserted into a plasmid pGEX-5X-1,at the downstream of the DNA sequence encoding Schistosoma japonicum glutathione S- transferase(GST),in the same reading frame with the gene of GST. A 60KD GST-NS53 fusion protein was expressed at 37℃ in a form of inclusion bodies amounting to 30 percent of total host protein whereas at 20℃ mainly in a form of soluble protein . The fusion protein was extracted and purified to homologue. The purified GST-NS53 fusion protein could be specifically recognized with either the sera from the patient infected by HGV or the antisera directed against GST.