Construction and identification of recombinant vaccine Bifidobacterium bifidum pGEX-Sj14-3-3 of Schistosoma japonicum

Q3 Medicine

中华地方病学杂志 Pub Date : 2011-07-20 DOI:10.3760/CMA.J.ISSN.1000-4955.2011.04.002

Ning Zhang, Li Wengui

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引用次数: 0

Abstract

Objective To construct and identify recombinant vaccine Bifwlobacterium bifidum(Bb)pGEX-Sj14-3-3 of Schistosoma japonicum(Sj). Methods Total RNA was extracted from adult Sj, antigen encoding gene Sj14-3-3 was amplified by RT-PCR and cloned into Escherichia coli (E. coli)-Bb shuttle expression vector pGEX-1λT to construct recombinant plasmid pGEX-Sj14-3-3. The recombinant plasmid was transformed into E. coli BL21 (DE3).The plasmid was extracted and identified by using BamH I and EcoR I. Then pGEX-Sjl4-3-3 was electroporated into Bb to construct recombinant Bb (pGEX-Sj14-3-3) vaccine. The extracted plasmid of the recombinant Bb (pGEX-Sj14-3-3) vaccine was identified by PCR, and the size of the products was compared with Sj14-3-3 gene of adult worms.Results Sj14-3-3 of 399 bp in length was amplified by RT-PCR. The products were digested by BamH I and EcoR I , and the fragments length of plasmid pGEX-Sj14-3-3 vector was 4947 bp, and of Sj 14-3-3 gene was 399 bp.The product of 399 bp Sj14-3-3 gene was also amplified by PCR from template of the extracted plasmid of the recombinant Bb(pGEX-Sj14-3-3 ) vaccine. The size of the product obtained was just the same as expected.Conclusion The recombinant Bb(pGEX-Sj14-3-3) vaccine of Sj is successfully constructed. Key words: Schistosoma japonicum; Bifidobacterium bifidum; Plasmids; Vaccines

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日本血吸虫两歧双歧杆菌重组疫苗pGEX-Sj14-3-3的构建与鉴定

目的构建并鉴定日本血吸虫双歧杆菌(Bb)重组疫苗pGEX-Sj14-3-3。方法提取成人Sj总RNA，通过RT-PCR扩增抗原编码基因Sj14-3-3，并将其克隆到大肠杆菌-Bb穿梭表达载体pGEX-1λT中，构建重组质粒pGEX-Sj14-3-3。将重组质粒转化大肠杆菌BL21 (DE3)。提取质粒，用BamHⅰ和EcoRⅰ进行鉴定，将pGEX-Sjl4-3-3电孔插入Bb中，构建重组Bb (pGEX-Sj14-3-3)疫苗。提取的重组Bb (pGEX-Sj14-3-3)疫苗质粒经PCR鉴定，产物大小与成虫Sj14-3-3基因进行比较。结果经RT-PCR扩增得到全长399 bp的Sj14-3-3。产物经BamH I和EcoR I酶切，质粒pGEX-Sj14-3-3载体片段长度为4947 bp, Sj14-3-3基因片段长度为399 bp。以重组Bb(pGEX-Sj14-3-3)疫苗质粒为模板，用PCR扩增出399bp的Sj14-3-3基因产物。得到的产品尺寸与预期的完全一致。结论成功构建了重组Bb(pGEX-Sj14-3-3)疫苗。关键词:日本血吸虫;双歧杆菌bifidum;质粒;疫苗

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来源期刊

中华地方病学杂志我国对人类健康危害特别严重的地方性疾病：克山病、大骨节病、碘缺乏病、地方性氟中毒、地方性砷中毒、鼠疫、布鲁氏菌病、寄生虫、新冠肺炎等疾病，同时还报道多发性自然疫源性疾病。

CiteScore

1.60

自引率

0.00%

发文量

8714

期刊介绍： The Chinese Journal of Endemiology covers predominantly endemic diseases threatening health of the people in the areas affected by the diseases including Keshan disease, Kaschin-Beck Disease, iodine deficiency disorders, endemic fluorosis, endemic arsenism, plague, epidemic hemorrhagic fever, brucellosis, parasite diseases and the diseases related to local natural and socioeconomic conditions; and reports researches in the basic science, etiology, epidemiology, clinical practice, control as well as multidisciplinary studies on the diseases.