摘要
目的从EST鉴定入手利用生物信息学方法筛选验证PCBP2基因内部UC.338基因座位附近的长非编码RNA。方法首先利用生物信息学方法筛选包含内含子转录区域的ESTs,经CPC分析和统计学比较,共筛选到4个(人、鼠各两个)可能的lncRNAs,用PCR方法进行鉴定,克隆到pGEM-T载体测序验证,并用PCR,real-time PCR的方法检测各ESTs在人源细胞系或小鼠不同组织,小鼠发育不同时间点大脑中的表达谱。结果筛选到4个ESTs中有3个ESTs均具有一定的非编码特征,并最终测序正确:1个人源,2个鼠源,并且其中有些lncRNAs具有一定的细胞、组织特异性,有些lncRNAs具有广谱表达模式。结论使用了一种新的寻找lncRNAs的方法,从EST鉴定入手,对现有数据库进行了挖掘。找到了可能在小鼠大脑发育过程中起作用的lncRNAs。
Objective Screening and identification of long noncoding RNAs in PCBP2 gene locus based on ESTs. To screen ESTs with introns transcription, analysis of CPC and statistical comparison, four ESTs were recognized as potential lncRNAs ( two of the four are human ESTs, the other two are mouse ESTs), those ESTs were identified with polymerase chain reaction(PCR) and then cloned into pGEM-T Vector-System for sequencing. Re suits The four ESTs are potential long noncoding RNAs, three of four ESTs were identified, their expression were different. Conclusions With a new method for searching lncRNAs based on ESTs, we identified some potential long noncoding RNAs, one of which may be a functional lncRNA influence mouse brain development.
出处
《基础医学与临床》
CSCD
北大核心
2013年第6期694-698,共5页
Basic and Clinical Medicine
基金
国家自然科学基金(31071203)