人源性蓝氏贾第鞭毛虫Elp3基因的克隆及序列分析

目的克隆蓝氏贾第鞭毛虫Elp3基因,并应用生物信息学方法进行分析。方法根据蓝氏贾第鞭毛虫Elp3已知基因序列的特点设计引物,利用PCR技术扩增获得Elp3的核苷酸序列,连接到pET28a载体并测定序列。应用生物信息学方法分析Elp3基因的序列同源性与结构特征。结果扩增片段长度为1767bp,测序结果与蓝氏贾第鞭毛虫WB株（GL50830）同源性100%。可构建生物进化树,进行同源结构建模发现,71-362aa具有一个保守的S-腺甙基蛋氨酸区域,458-584aa具有组蛋白乙酰基转移酶区域。结论成功克隆了蓝氏贾第鞭毛虫Elp3基因,生物信息学分析发现,其在进化上与其它物种不属同一起源,具有SAM和HAT的结构和功能,这些发现为进一步研究其生物学功能提供了依据和线索。

Cloning and sequence analysis of the gene encoding Elp3 of Giardia lamblia isolated from human

Objective To obtain and analyze the sequence of SUCH/89/BTMRI/2（C2） Elp3 gene.Methods Primers were designed according to the published G.lamblia Elp3 gene sequence.Elp3 gene was amplified from G.lamblia C2 genomic DNA by PCR.PCR product was then inserted into pET28a Simple Vector and transformed into E.coli JM109.Positive clone was identified.Bioinformatics technique was used to analyze the gene sequence homology and structural feature of Elp3.Results A 1 767 bp long fragment of the G.lamblia Elp3 gene was successfully amplified and cloned.The obtained gene fragment was highly homologous to G.lamblia（GL50803）.Phylogenetic tree of Elp3 was then constructed,and the tertiary structure prediction showed that Elp3 had a s-adenosylmethionine region in 71-362aa and a histone acetyltransferase region in 458-584aa.Conclusion G.lamblia Elp3 gene was cloned successfully and analyzed by bioinformatics technique.The origin of this gene is not similar with other species.The gene has the structure and function of SAM and HAT region.Our results provide some evidences and clues for the further study on biological functions of G.lamblia Elp3 gene.