摘要
为进一步确定柔嫩艾美耳球虫(E.tenella)端粒DNA的重复序列,为下一步端粒酶活性检测奠定基础,根据已克隆发表的E.tenella端粒DNA重复序列信息,设计了由4个端粒重复序列串联的寡聚核苷酸探针(TTTAGGG)4,并以生物素地高辛标记。将该探针与经BAL31-EcoRⅠ酶切后的E.tenella基因组DNA进行Southern印迹杂交分析。结果显示:E.tenella基因组DNA与探针杂交获得了清晰的杂交条带,随BAL31酶切时间的延长,杂交信号逐渐减弱,进一步证明了E.tenella端粒DNA重复序列为5-′TTTAGGG-3′,且此重复序列在E.tenella染色体的末端。
In order to identify E.tenella telomeric repetitive DNA sequences and detect the activity of telomerase,an oligonucleotide probe composed of（TTTAGGG）4 telomeres was designed according to the E.tenella telomeric repetitive DNA sequence（5′-TTTAGGG-3′） which had been previously cloned in our laboratory and labeled by digoxin.The probe was hybridized with the products of E.tenella genomic DNA digested with BAL31 and EcoRⅠand then analyzed by Southern blotting.The results indicated that the hybridization of the probe and the E.tenella genomic DNA produced clear bands.With the delay of BAL31 digestion,hybridization signal decreased gradually.It further proved that E.tenella telomeric repetitive DNA sequences were 5′-TTTAGGG-3′and located in the terminal region of E.tenella chromosome.
出处
《吉林农业大学学报》
CAS
CSCD
北大核心
2010年第2期205-208,共4页
Journal of Jilin Agricultural University
基金
国家自然科学基金项目(30671580
30170696)
