| 弓形虫ROP2基因的克隆及在大肠杆菌中的表达 |
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| 引用本文： | 吴昆,陈晓光,支国舟. 弓形虫ROP2基因的克隆及在大肠杆菌中的表达[J]. 寄生虫与医学昆虫学报, 2004, 11(3): 135-138 |
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| 作者姓名： | 吴昆 陈晓光 支国舟 |
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| 作者单位： | 第一军医大学寄生虫学教研室,广州,510515;第一军医大学寄生虫学教研室,广州,510515;第一军医大学寄生虫学教研室,广州,510515 |
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| 基金项目： | 广东省科技计划项目 (A10 90 2 0 5 ) |
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| 摘 要： | 目的 :构建弓形虫棒状体蛋白 (ROP2 )基因重组质粒并在大肠杆菌中表达 ,用于筛选弓形虫新的诊断抗原和疫苗分子。方法 :根据ROP2基因已知序列 ,设计合成一对引物 ,用PCR方法从弓形虫RH株基因组DNA中扩增出ROP2基因片段 ,再克隆到pGEX 4T 1质粒 ,重组质粒经酶切及PCR鉴定 ,而后进行测序 ;重组质粒在大肠杆菌BL2 1 (DE3)中进行ITPG诱导表达 ,表达产物经SDS PAGE及WesternBlot分析。结果 :ROP2基因PCR产物大小与预期相符 ,约 1 0 4 3bp ,重组质粒经酶切及PCR鉴定表明获得正确重组子 ,测序结果与已知序列基本吻合 ;SDS PAGE及免疫印迹显示ROP2融合蛋白表观分子量约为 5 5kDa ,表达产物约占菌体总蛋白1 7% ,具有一定的免疫反应性。结论 :克隆并表达了弓形虫ROP2基因 ,为下一步弓形虫诊断及疫苗研究奠定了基础
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| 关 键 词： | 刚地弓形虫 ROP 2 基因克隆 融合表达 |
| 修稿时间： | 2004-03-23 |
CLONING AND EXPRESSION OF TOXOPLASMA GONDII ROP2 GENE IN E.COLI |
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| WU Kun CHEN Xiao Guang ZHI Guo Zhou. CLONING AND EXPRESSION OF TOXOPLASMA GONDII ROP2 GENE IN E.COLI[J]. Acta Parasitologica et Medica Entomologica Sinica, 2004, 11(3): 135-138 |
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| Authors: | WU Kun CHEN Xiao Guang ZHI Guo Zhou |
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| Abstract: | Objective:To construct a recombinant plasmid containing rhoptry protein 2(ROP2)gene of Toxoplasma gondii and express it in E.coli for selection of diagnostic antigen and vaccine candidate.Method:The truncated ROP2 gene was amplified from the genomic DNA of Toxoplasma gondii RH strain and cloned into plasmid pGEX 4T 1.The expression of the recombinant pGEX 4T 1 ROP2 was induced by IPTG in BL21(DE3).The expressed proteins were analysed by SDS|PAGE and Western blot.Result:The amplified ROP2 gene was about 1043 bp and confirmed by DNA sequencing.SDS PAGE and Western blot showed a position band around 55 kDa in size.Conclusion:The truncated ROP2 gene of Toxoplasma gondii has been successfully cloned and expressed in BL21(DE3)cells. |
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| Keywords: | Toxoplasma gondii Rhoptry protein 2 Gene clone Protein expression |
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