贾第虫核糖核酸酶PRNA转录本3′端序列及其存在形式  

The 3′-terminal sequence of P RNA transcript and its form in Giardia

作　　者：高珺珊[1] 吴威[1] 宫鹏涛[1] 李建华[1] 杨举[1] 李赫[1] 张国才[1] 张西臣[1] 

机构地区：[1]吉林大学动物医学学院,吉林长春130062

出　　处：《中国生物制品学杂志》2013年第8期1122-1125,共4页Chinese Journal of Biologicals

基　　金：国家自然科学基金项目(30970322)

摘　　要：目的确定贾第虫滋养体内核糖核酸酶P(RNase P)RNA 3′端序列及其存在形式。方法提取贾第虫滋养体总RNA,大肠埃希菌(E.coli)Poly(A)聚合酶加polyA尾后,进行反转录,扩增出加polyA尾后的cDNA,经PCR及测序进行鉴定,确定其3′端序列。应用实时荧光定量PCR(RT-qPCR)分别检测RNase P-GLsR15共转录体与RNase PRNA成熟体的总表达量,两者之差即为RNase P RNA成熟体的表达量,确定贾第虫RNase P RNA的存在形式。结果 RNase P RNA 3′cDNA大小约300 nt,3′端序列与GLsR15 3′端序列一致;RNase P-GLsR15共转录体和RNaseP RNA成熟体的总表达量与RNase P-GLsR15共转录体表达量差异无统计学意义(P>0.05)。结论已成功克隆了RNase P RNA 3′端序列,证实RNase P RNA和GlsR15的共转录体即为RNase P RNA成熟体的存在形式。Objective To determine the 3′-terminal sequence of P RNA transcript and its form in Giardia.Methods Total RNA of the trophozoites of Giardia were extracted and polyadenylated,then reversely transcribed for 3′RACE with specific and adapter primers.The obtained cDNA with a poly A tail was identified by PCR and sequencing,of which the 3′-terminus was determined.The total expression levels of co-transcripted RNase P-GLsR15 RNA and matured RNase P RNA were determined by real-time fluorescent quantitative PCR（Rt-qPCR）.Results The length of RNase P RNA 3′ cDNA was about 300 nt,of which the sequence at 3′-terminus was consistent with that of GLsR15.No significant differences were observed in total expression levels of co-transcripted RNase P-GLsR15 RNA and matured RNase P RNA（P 0.05）.Conclusion The 3′-terminal sequence of RNase P RNA was successfully cloned,which confirmed that the cotranscripted RNase P-GLsR15 RNA was the existing form of matured RNase P RNA.

关 键 词：贾第虫属 核糖核酸酶 3′RACE 共转录体 

分 类 号：R382.9[医药卫生—医学寄生虫学]