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目的获取刚地弓形虫(Toxoplasma gondii)RH株表面抗原SAG3目的基因片段,对其结构和功能进行生物信息学分析,预测其编码蛋白作为候选抗原基因的可行性。方法提取弓形虫的基因组DNA,自行设计一对寡核苷酸引物,PCR体外扩增SAG3基因,用生物信息学方法对SAG3蛋白的理化性质、结构和功能进行预测。结果扩增出的基因片段约1174bp,测序后鉴定其为弓形虫RH株SAG3基因片段。预测SAG3蛋白相对分子质量约为41785.2,能形成两个功能结构域,氨基酸序列的前38(或39)位为信号肽序列。通过多种预测方法分析SAG3氨基酸序列抗原表位可能位于第10、50、80、95、160、180、220、250、300、330和360位点内或附近。结论成功获得SAG3基因,为深入研究该基因结构奠定了基础。
OBJECTIVE: To obtain the SAG3 gene fragment of Toxoplasma gondii RH strain surface antigen and analyze the structure and function of SAG3 gene by bioinformatics analysis, and to predict the feasibility of its encoded protein as a candidate antigen gene. Methods Toxoplasma gondii genomic DNA was extracted and a pair of oligonucleotide primers was designed. SAG3 gene was amplified by PCR in vitro. The physical and chemical properties, structure and function of SAG3 protein were predicted by bioinformatics methods. Results The amplified fragment was about 1174bp. After sequencing, it was identified as the SAG3 gene fragment of T. gondii RH strain. The relative molecular mass of SAG3 protein is predicted to be about 41785.2, forming two functional domains. The first 38 (or 39) of the amino acid sequence is the signal peptide sequence. Analysis of SAG3 amino acid sequence epitopes by a variety of prediction methods may lie within or near the 10th, 50th, 80th, 95th, 160th, 180th, 220th, 250th, 300th, 330th, and 360th sites. Conclusion The SAG3 gene was successfully obtained and laid the foundation for further study on the gene structure.