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目的比较弓形虫强毒株RH株、桂弓株和弱毒株B36株致密颗粒蛋白1(GRA1)基因片段的异同。方法分别从接种RH株、桂弓株、B36株弓形虫的小鼠腹水中收集、纯化虫体,提取基因组DNA,PCR扩增3个虫株的致密颗粒蛋白1(GRA1)目的基因片段;构建pMD-18-T/GRA1重组质粒,PCR和双酶切鉴定其准确性;测序并用NCBI中Blast软件和MEGA3.1软件分析比较3种不同毒力株GRA1基因的异同。结果获得3株弓形虫的GRA1目的基因片段并构建了pMD-18-T/GRA1重组质粒,经鉴定目的基因片段大小与预期理论值相符。测序分析3不同毒力株GRA1基因片段相似为100%。结论弓形虫强毒株RH株、桂弓株和弱毒株B36株的GRA1基因片段极其相似,且高度保守。因此认为弓形虫GRA1基因具有较高应用价值。
Objective To compare the similarities and differences between the GRA1 gene fragments of RH strain of Toxoplasma gondii, Gui bow strain and attenuated strain B36. Methods The worms were collected and purified from the ascites of mice inoculated with Rh strain, Gui Gong strain and Toxoplasma gondii strain B36 respectively. The genomic DNA was extracted and the GRA1 gene fragments of three strains were amplified by PCR. pMD-18-T / GRA1 recombinant plasmid, PCR and double digestion to identify its accuracy; sequencing and NCBI Blast software and MEGA3.1 software analysis of three different virulence strains GRA1 gene similarities and differences. Results Three GRA1 gene fragments of Toxoplasma gondii were obtained and the recombinant plasmid pMD-18-T / GRA1 was constructed. The size of the identified gene fragment was consistent with the expected theoretical value. Sequencing Analysis 3 The GRA1 gene fragments of different virulent strains were similar to 100%. Conclusion The GRA1 gene fragments of T. gondii RH strain, Gui Gong strain and attenuated strain B36 strain are very similar and highly conserved. So that Toxoplasma GRA1 gene has a high value.