日本血吸虫重组两歧双歧杆菌疫苗的构建及其表达  

Construction and expression of a recombinant Bifidobacterium bifidum vaccine of Schistosoma japonicum

作　　者：罗广旭[1] 李文桂[1] 覃婷[1] 

机构地区：[1]重庆医科大学附属第一医院传染病寄生虫病研究所,重庆400016

出　　处：《中华地方病学杂志》2017年第4期257-260,共4页Chinese Journal of Endemiology

基　　金：重庆市科委地方病重大专项基金（2008AB5055、2008AB5008、2008AB5054）

摘　　要：目的构建及鉴定日本血吸虫（Sj）重组两歧双歧杆菌（Bb）疫苗[Bb（pGEX—Sj26GST—Sj14-3-3）]，并研究融合基因Sj26GST—Sj14—3—3在Bb中的表达情况。方法将重组质粒pGEX—Sj26GST—Sj14-3-3电转化至Bb中，构建重组Bb（pGEX—SjZ6GST-Sj14-3—3）疫苗，用异丙基硫代-β-D-半乳糖苷（IPTG）诱导表达。采用双酶切和聚合酶链式反应（PCR）鉴定重组Bb（pGEX—Sj26GST-Sj14—3—3）疫苗，采用十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（SDS—PAGE）和蛋白免疫印迹法对表达产物进行鉴定。结果双酶切及PCR证实重组质粒pGEX—SjZ6GST—Si14—3—3成功转入Bb中。SDS—PAGE鉴定结果显示，诱导表达产物是相对分子质量约为67×10^3的重组蛋白，与预期结果相符。蛋13免疫印迹法鉴定结果显示，重组蛋白能被Sj感染的兔血清所识别。结论成功构建Sj重组Bb（pGEX—Sj26GST-Sj14-3—3）疫苗，Rb可表达具有特异抗原性的Sj26GST—Sj14—3．3重组蛋白。Objective To construct a recombinant Bifidobacterium bifidum （Bb） vaccine [Bb （pGEX- Sj26GST-Sj 14-3-3）] of Schistosomajaponicum （Sj） and analyze the expression of the fusion gene Sj26GST-Sj 14-3-3 of Sj in Bb. Methods The recombinant plasmid pGEX-Sj26GST-Sj14-3-3 was electroporated into Bb to construct a recombinant Bb （pGEX-Sj26GST-Sj14-3-3） vaccine. After induction with isopropyl-f3-D-thiogalactoside （IPTG）, double restriction enzymes digestion and polymerase chain reaction （PCR） were used to identify the recombinant Bb （pGEX-Sj26GST-Sj14-3-3）, expression of the recombinant protein was analyzed and identified by sodium dodecyl sulfonate-polyacrylamide gel electrophoresis （SDS-PAGE） and Western blotting. Results The recombinant plasmid pGEX-Sj26GST-Sj14-3-3 was successfully transformed into Bb identified by double restriction enzymes digestion and PCR. SDS-PAGE analysis showed that the relative molecular mass of the expressed recombinant protein was approximately 67 × 10^3. The expressed protein could be recognized by the immune sera from rabbits infected with Sj by Western blotting. Conclusions The recombinant Bb （pGEX-Sj26GST-Sj14-3-3） vaccine of Sj is successfully constructed. The fusion gene Sj26GST-Sj14-3-3 can be expressed in recombinant Bb and the expressed target protein shows specific antigenicity.

关 键 词：血吸虫 日本 融合基因 两歧双歧杆菌 疫苗 

分 类 号：R392[医药卫生—免疫学]