中国寄生虫学与寄生虫病杂志
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中国寄生虫学与寄生虫病杂志 2008, Vol. 26 Issue (2) :127    DOI:
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日本血吸虫SjPP基因重组杆状病毒的构建和在昆虫细胞中的表达
姚利晓1,3,陶丽红1,杨冠珍2,吴祥甫2,蔡幼民1,林矫矫1 *
1 中国农业科学院上海兽医研究所，农业部动物寄生虫学重点开放实验室，上海动物生物技术研究中心，上海 200232； 2 中国科学院上海生命科学研究院上海生化细胞所，上海 200231；3 西南大学柑桔研究所，重庆 400712
Construction and Expression of Recombinant Baculovirus with Schistosoma japonicum SjPP Gene
YAO Li-xiao1，3， TAO Li-hong1， YANG Guan-zhen2， WU Xiang-fu2， CAI You-min1， LIN Jiao-jiao1 *
1 Shanghai Research Center for Animal Biotechnology/Key Laboratory of Animal Parasitology of the Ministry of Agriculture/ Shanghai Institute of Veterinary Research，Chinese Academy of Agricultural Sciences，Shanghai 200232，China；2 Institute of Biochemistry and Cell Biology，Shanghai Institutes for Biological Sciences，Chinese Academy of Sciences，Shanghai 200231，China；3 Citrus Research Institute, Southwest University, Chongqing 400712，China
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摘要  目的 在昆虫细胞中表达获得可溶性的日本血吸虫SjPP重组蛋白。 方法 提取日本血吸虫成虫总RNA，通过RT-PCR扩增出SjPP基因的编码区序列，将其定向克隆到供体质粒pFastBac HT-B中，构建重组杆状病毒供体质粒pFastBac-SjPP，转化入大肠埃希菌DH10Bac进行转座，提取重组杆粒Bacmid-SjPP，用阳离子脂质体法转染粉纹夜蛾（TN5B1-4）细胞。利用十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（SDS-PAGE）和蛋白质印迹（Western blotting）分析重组蛋白表达情况。 结果 构建了含日本血吸虫SjPP基因的重组杆粒Bacmid-SjPP，转染TN5B1-4细胞后，获得有感染力的重组杆状病毒，并在TN5B1-4细胞中成功表达SjPP蛋白。该重组蛋白可被抗SjPP蛋白的兔血清识别。 结论 成功构建SjPP基因重组杆状病毒，并在TN5B1-4细胞中表达，该蛋白具有良好的抗原性。
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姚利晓
陶丽红
杨冠珍
吴祥甫
蔡幼民
林矫矫
关键词： 日本血吸虫   SjPP   Bac-to-Bac杆状病毒表达系统     
Abstract： Objective To express the soluble recombinant Schistosoma japonicum SjPP proteins in TN5B1-4 cells. Methods The total RNA was extracted from adult worms of Schistosoma japonicum. The whole coding sequence of SjPP gene was synthesized by RT-PCR and cloned into donor plasmid. The recombinant donor pFastBac-SjPP was transformed into E.coli DH10Bac forming Bacmid-SjPP which was transfected into insect cell with cational lipofectin. The fusion protein SjPP was analyzed with SDS-PAGE and Western blotting. Results The infective recombinant baculovirus Bacmid-SjPP was obtained and SjPP protein was expressed in insect cells. Conclusion The recombinant protein SjPP has been expressed in insect TN5B1-4 cells with proper antigenicity.
Keywords： Schistosoma japonicum   SjPP   Bac-to-Bac Baculovirus Expression Vector System     
Received 1900-01-01; published 2008-04-30
Corresponding Authors: 林矫矫   
引用本文:   
姚利晓;陶丽红;杨冠珍;吴祥甫;蔡幼民;林矫矫.日本血吸虫SjPP基因重组杆状病毒的构建和在昆虫细胞中的表达[J]  中国寄生虫学与寄生虫病杂志, 2008,V26(2): 127
YAOLi-xiao;TAOLi-hong;YANGGuan-zhen;WUXiang-fu;CAIYou-min;LINJiao-jiao*.Construction and Expression of Recombinant Baculovirus with Schistosoma japonicum SjPP Gene[J]  , 2008,V26(2):127
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