C2株蓝氏贾第鞭毛虫组蛋白H4基因的克隆与同源分析.DOCVIP

C2株蓝氏贾第鞭毛虫组蛋白H4基因的克隆与同源性分析 余源 李冀 沈海娥 李明 何丽萍 田喜凤* 河北联合大学生命科学学院，唐山063000 摘要 目的 从C2株蓝氏贾第鞭毛虫基因组中获得组蛋白H4基因序列并进行同源比对。方法 应用PCR技术从蓝氏贾第鞭毛虫基因组中获得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，与美国WB株蓝氏贾第鞭毛虫及其他物种H4组蛋白基因和蛋白序列进行同源分析。结果 成功克隆并得到C2株蓝氏贾第鞭毛虫H4组蛋白基因序列，同源比对结果显示其基因序列与美国WB株完全一致，进化树分析结果表明贾第虫组蛋白H4基因在进化上较为独立，与其他物种存在较大分歧。结论 蓝氏贾第鞭毛虫与其他现存真核生物亲缘关系较为疏远，为进一步研究蓝氏贾第鞭毛虫的生物进化地位提供实验资料。 关键词 C2株蓝氏贾第鞭毛虫 组蛋白 同源分析 Cloning and Homologous alignment of H4 histone gene in C2 Giardia lambliaYuyuan liji shenhaie tianxifeng Colledge of life science of Hebei United Uinion 063000 Abstract: Objective To obtain histone H4 gene sequence from C2 Giardia lamblia genome DNA and homologous alignment. Methods The histone H4 gene sequence of Giardia lamblia was amplified from genomic DNA by PCR and restructured into pGM-T vector. Transforming TOP10 competent bacteria cells with recombinant plasmid DNA. Positive cloning was chosen and the sequence was analysed. The coding sequence of H4 was aligned with WB Giardia lamblia histone H4 gene sequence and the other organisms. Results The histone H4 gene sequence of C2 Giardia lamblia was successfully cloned and obtained, Homologous alignment results shown that histone H4 gene sequence of C2 Giardia lamblia was the same with histone H4 gene sequence of WB Giardia lamblia, the phylogenetic tree analysis results shown that histone H4 gene of C2 Giardia lamblia was independent in biological evolution, there was a big difference with other species. Conclusions The genetic relationship was distant between Giardia lamblia and other existing eukaryotes, which provided valuable experimental data for further understanding the biological evolution orientation of Giardia lamblia. 国家自然科学基金（No.和唐山市科技支持计划项目（N0:-33）联合资助 Supported by: Grant No.from the National Natural Science Foundation of China; Scientific and technological support projects from Tangshan, China Grant No.-33 **通讯作者: 田喜凤E-mail tstianxifeng@163.com 作者简介：余源，