CAJ | 학술논문

目的：构建猪带绦虫重组BCG‐TSOL18疫苗，研究TSOL18基因在BCG中的表达情况。方法通过酶切的方法从重组质粒pGEX‐TSOL18获取猪带绦虫TSOL18基因，将其定向克隆到大肠杆菌‐分枝杆菌穿梭表达质粒pMV261中，构建猪带绦虫重组质粒pMV261‐TSOL18，进行酶切、PCR和测序鉴定；再将其电穿孔转化入BCG ，构建猪带绦虫重组BCG‐TSOL18疫苗，进行PCR鉴定；SDS‐PAGE和Western blot分析 TSOL18基因在BCG中的表达情况。结果通过酶切成功获得了393 bp的TSOL18基因片段；酶切、PCR和测序鉴定证明成功构建了猪带绦虫重组质粒pMV261‐TSOL18；PCR鉴定证实猪带绦虫重组质粒pMV261‐TSOL18成功转入BCG中，提示猪带绦虫重组BCG‐TSOL18疫苗构建成功；SDS‐PAGE分析发现在相对分子质量（M r）约为14．7 kD处有明显的TSOL18目的蛋白条带，Western blot证实表达的TSOL18目的蛋白能被兔抗血清和囊虫病猪血清所识别。结论成功构建了猪带绦虫重组BCG‐TSOL18疫苗，TSOL18基因能够在BCG中成功表达，表达的TSOL18重组蛋白具有特异的抗原性，为该疫苗的进一步研究奠定了基础。
목적：구건저대조충중조BCG‐TSOL18역묘，연구TSOL18기인재BCG중적표체정황。방법통과매절적방법종중조질립pGEX‐TSOL18획취저대조충TSOL18기인，장기정향극륭도대장간균‐분지간균천사표체질립pMV261중，구건저대조충중조질립pMV261‐TSOL18，진행매절、PCR화측서감정；재장기전천공전화입BCG ，구건저대조충중조BCG‐TSOL18역묘，진행PCR감정；SDS‐PAGE화Western blot분석 TSOL18기인재BCG중적표체정황。결과통과매절성공획득료393 bp적TSOL18기인편단；매절、PCR화측서감정증명성공구건료저대조충중조질립pMV261‐TSOL18；PCR감정증실저대조충중조질립pMV261‐TSOL18성공전입BCG중，제시저대조충중조BCG‐TSOL18역묘구건성공；SDS‐PAGE분석발현재상대분자질량（M r）약위14．7 kD처유명현적TSOL18목적단백조대，Western blot증실표체적TSOL18목적단백능피토항혈청화낭충병저혈청소식별。결론성공구건료저대조충중조BCG‐TSOL18역묘，TSOL18기인능구재BCG중성공표체，표체적TSOL18중조단백구유특이적항원성，위해역묘적진일보연구전정료기출。
We constructed a recombinant Bacillus Calmette‐Guerin(BCG)‐TSOL18 vaccine of Taenia solium and observed the expression of the TSOL18 gene in BCG .The TSOL18 gene of Taenia solium was obtained from the recombinant plasmid pGEX‐TSOL18 by digestion method and cloned into Escherichia coli (E .coli)‐mycobacterium shuttle plasmid pMV261 to con‐struct the recombinant plasmid pMV261‐TSOL18 of Taenia solium ,and the recombinant plasmid was identified by restriction enzyme digestion ,PCR and DNA sequencing .Then ,the recombinant plasmid was transformed into BCG by electroporation to construct the recombinant BCG‐TSOL18 vaccine of Taenia solium ,and the vaccine was identified by PCR .The expression of the TSOL18 gene in BCG was identified by SDS‐PAGE and Western blot .The 393 bp TSOL18 gene fragment was successfully obtained by restriction enzyme digestion .Restriction enzyme digestion ,PCR and DNA sequencing suggested that the recombi‐nant plasmid pMV261‐TSOL18 of Taenia solium was successfully constructed .PCR confirmed that the recombinant plasmid pMV261‐TSOL18 of Taenia solium was successfully transformed into BCG ,suggesting that the recombinant BCG‐TSOL18 vaccine of Taenia solium was successfully constructed .SDS‐PAGE showed that the relative molecular mass (Mr) of TSOL18 target protein was approximately 14 .7 kD .Results of western blot showed the TSOL18 target protein could be recognized by rabbit antiserum or cysticercosis swine serum .The recombinant BCG‐TSOL18 vaccine of Taenia solium was successfully con‐structed .The TSOL18 gene of Taeniasolium was successfully expressed in BCG and the expressed TSOL18 recombinant pro‐tein had specific antigenicity .This result would lay a foundation for further study of the vaccine .