CAJ | 학술논문

　　本研究旨在用杆状病毒/昆虫细胞表达系统表达日本血吸虫热休克70蛋白(SjHSP70)，获得重组的 rSjHSP70蛋白。利用 PCR 技术从日本血吸虫 cDNA 文库中克隆 SjHSP70全长序列；亚克隆到 pFastBacHTA 中，构建重组杆状病毒转移载体pFastBacHTA-SjHSP70，导入 DH10Bac 感受态细胞，得到重组穿梭质粒 reBacmid-SjHSP70，再转染到 Sf9昆虫细胞制备重组病毒；将重组病毒感染细胞，用间接免疫荧光技术鉴定重组蛋白的表达；His 柱亲和层析法纯化目的蛋白；Western blot 技术对纯化蛋白进行抗原性鉴定。SjHSP70基因全长序列1962 bp，在 Sf9昆虫细胞中获得高效表达；His 柱亲和层析法纯化表达的蛋白可被日本血吸虫感染小鼠的阳性血清识别。用杆状病毒昆虫细胞表达系统成功表达了 SjHSP70，为深入研究 SjHSP70分子的免疫生物学功能提供了材料。
　　본연구지재용간상병독/곤충세포표체계통표체일본혈흡충열휴극70단백(SjHSP70)，획득중조적 rSjHSP70단백。이용 PCR 기술종일본혈흡충 cDNA 문고중극륭 SjHSP70전장서렬；아극륭도 pFastBacHTA 중，구건중조간상병독전이재체pFastBacHTA-SjHSP70，도입 DH10Bac 감수태세포，득도중조천사질립 reBacmid-SjHSP70，재전염도 Sf9곤충세포제비중조병독；장중조병독감염세포，용간접면역형광기술감정중조단백적표체；His 주친화층석법순화목적단백；Western blot 기술대순화단백진행항원성감정。SjHSP70기인전장서렬1962 bp，재 Sf9곤충세포중획득고효표체；His 주친화층석법순화표체적단백가피일본혈흡충감염소서적양성혈청식별。용간상병독곤충세포표체계통성공표체료 SjHSP70，위심입연구 SjHSP70분자적면역생물학공능제공료재료。
The objective of the present study was to express recombinant SjHSP70 in insect cells Sf9. The SjHSP70 gene of 1962 bp was amplified by PCR and used to construct vector pFastBacHTA-SjHSP70. Then, reBacmid-SjHSP70 was transformed into competent cells. The recombinant shuttle plasmid reBacmid-SjHSP70 was obtained from the transformed DH10Bac cells and then transfected into insect cells Sf9. The recombinant protein SjHSP70 expressed in infected cells was visualized in IFA and purified with His column affinity chromatography. Western blot demonstrated the ecombinant SjHSP70 was recognized by antiserum from mice infected with Schistosoma japonicum. The availability of recombinant SjHSP70 provided materials for further exploring its immunobiological function.