摘要
目的从C2株蓝氏贾第鞭毛虫(简称贾第虫)基因组中获得组蛋白H4基因序列并进行同源比对。方法采用组织/细胞基因组DNA提取试剂盒提取贾第虫的基因组DNA,利用Primer Premier 5软件设计特异性引物,应用PCR技术从贾第虫基因组中获得H4组蛋白基因,构建pGM-T重组载体,转化TOP10感受态细菌,挑选阳性克隆并进行序列分析,分析结果与美国WB株贾第虫及其他模式生物H4组蛋白基因和蛋白序列分别进行同源比对。结果成功克隆并得到C2株贾第虫H4组蛋白基因序列,基因片段大小为310bp,同源比对结果显示其基因序列与美国WB株完全一致,进化树分析表明C2株贾第虫组蛋白H4基因在生物进化上较为独立,与其他物种存在较大分歧。结论C2株贾第虫与其他现存真核生物亲缘关系较为疏远,为进一步研究贾第虫的生物进化地位提供了实验资料。
Objective To obtain the sequence of the histone H4 gene from genomic DNA of the Giardia lamblia C2 strain and compare its homology. Methods Genomie DNA of G. lamblia was extracted using a tissue/cell genomic DNA extraction kit, and Primer Premier 5 was used to design specific primers. The sequence of the histone H4 gene of G. lamblia was amplified from genomic DNA using PCR and ligated into a pGM T vector. The vector was transformed into TOPI 0 competent cells with recombinant plasmid DNA. The positive clone was sequenced. The sequence of H4 was compared to that of the H4 gene of the G. lamblia WB strain and other model organisms. Results The sequence of the histone H4 gene of the G. lamblia C2 strain was successfully cloned and analyzed. The gene fragment was 310 bp. Comparison of homology indicated that the sequence of the histone H4 gene of the G. lamblia C2 strain was homologous with the sequence of the histone H4 gene of the G. lamblia WB strain. Analysis of the phylogenetic tree indicated that the his tone H4 gene of the G. lamblia C2 strain had evolved rather independently, and it differed substantially from other species. Conclusion The G. lamblia C2 strain has a distant genetic relationship to other known eukaryotes. This provided valuable experimental data for further study of the evolutionary position of （5. lamblia.
出处
《中国病原生物学杂志》
CSCD
北大核心
2013年第9期807-809,823,共4页
Journal of Pathogen Biology
基金
国家自然科学基金项目(No.30970313)
唐山市科技支持计划项目(No.12140209A-33)
