摘要
目的 :构建弓形虫致密颗粒抗原 1 (GRA1 )基因真核表达质粒。方法 :根据 GRA1基因已知序列 ,设计一对引物。用 PCR技术从弓形虫 RH株基因组 DNA中扩增 GRA1基因片段 ,插入 pc DNA3质粒 ,转化大肠杆菌 DH5 α感受态细胞 ,经酶切及 PCR鉴定、测序。结果 :从弓形虫 RH株基因组中扩增出 775 bp的 GRA1基因 ,测序显示该基因存在一 1 35 bp的插入序列 ,使得所得 PCR产物分子量大于预期值 (6 2 8bp)。该插入序列造成GRA1基因移码突变。结论 :首次报道了变异的弓形虫 RH株
Objective To construt a recombinant eukaryotic expressing plasmid containing dense granules antigen (GRA1) gene of Toxoplasma gondii . Methods A pair of primers were designed for PCR according to the known sequence of GRA1 gene.The full fragment of GRA1 gene obtained by PCR amplification from genomic DNA of RH strain of Toxoplasma gondii was cloned into plasmid pcDNA3 orientatedly.The constructed recombinant plasmid pcDNA3 GRA1 was transferred into E.coli DH5α.The transformatants were screened and identified by restriction endonuclease digestion and PCR. The nucleotide sequences of the cloned genes were determined. Results A 775 bp GRA1 gene PCR product was obtained from genomic DNA of the RH strain of Toxoplasma gondii ,in which a 135 bp inserted sequence was found.Therefore the PCR product was longer than expected (628 bp).The frameshift mutation for GRA1 gene resulted from the insertion of exogenous sequence. Conclusion The inserted variation of GRA1 gene of the RH strain of Toxoplasma gondii is reported and its recombinant expressing plasmid is construted.
出处
《吉林大学学报（医学版）》
CAS
CSCD
北大核心
2004年第3期369-371,共3页
Journal of Jilin University：Medicine Edition
基金
吉林省科技厅资助课题 (1 9980 334)
