犬贾第虫端粒酶催化亚基基因的克隆及序列分析

Clone and sequence analysis of telomerase reverse transcriptase gene from Giardia canis

目的对犬贾第虫端粒酶催化亚基(GcTERT)基因进行克隆及序列分析。方法根据人源贾第虫的端粒酶催化亚基TERT基因(AF195121)设计1对引物,以犬贾第虫滋养体总核酸为模板,扩增犬贾第虫端粒酶催化亚基(GcTERT)基因,PCR产物连接到PGEM-T载体并转化入DH5α感受态菌进行测序,通过BLAST对GenBank进行同源性搜索,并用分子生物学软件进行分析。结果测得犬贾第虫端粒酶催化亚基(GcTERT)基因全长2 883 bp,该基因序列与人源贾第虫的端粒酶催化亚基基因同源性为99%,氨基酸的同源性也为99%。结论成功扩增了犬贾第虫端粒酶催化亚基(GcTERT)基因,为进一步研究该基因的功能及筛选抗犬贾第虫药物靶标奠定了基础。

Objective To clone and analyze the gene of the telomerase reverse transcriptase from Giardia canis （GeTERT）. Methods According to the published nucleotide sequence of TERT of Giardia （AF195121）, a pair of primers were designed and total nucleic acids from trophozoites of G. canis were used as template for PCR. The PCR products were linked into pGEM-T vector, then cloned, sequenced and analyzed. Results The sequence of GcTERT was 2 883 bp and the homology of GcTERT to that of sequences （AF195121） was 99 % in the nucleotide, translating into a protein of 960 amino acids（aa）, the GcT ERT protein shared 99 % aa identity with TERT of Giardia （AF195121）. Conclusion The gene of GcTERT is successfully cloned, which is a basis to study on the function of GcTERT.