Analysis of the 3′ UTR of the ART3 and ART4 gene by 3′ inverse RACE-PCR

被引:7
作者
Friedrich, M [1 ]
Grahnert, A [1 ]
Hauschildt, S [1 ]
机构
[1] Univ Leipzig, Dept Immunobiol, Inst Biol 2, D-04103 Leipzig, Germany
来源
DNA SEQUENCE | 2005年 / 16卷 / 01期
关键词
3 ' untranslated region; RACE-PCR; inverse PCR; ADP-ribosyltransferase;
D O I
10.1080/10425170400025307
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
3' Rapid amplification of cDNA ends (3' RACE) is a polymerase chain reaction (PCR) based technique which has been developed to analyse 30 ends of partially known cDNA sequences. To improve the effectiveness of the technique, many investigators have modified the RACE protocol. Here, we describe an alternative procedure for analysing 30 mRNA ends which is based on DNA ligase-mediated self circularization and inverse PCR. This technique is simple and characterized by the exclusive use of gene-specific primers and the absence of unspecific adaptor sequences to obtain highly specific PCR products. We applied the method to analyze the 3' UTR of human mono-ADP- ribosyltransferase (ART) 3 mRNA in testis and heart muscle and of ART4 mRNA in HEL cells. The obtained sequences of ART3 and ART4 mRNA corresponded to data base entries of the respective mRNAs. No adenylate/uridylate-rich elements (AREs) were found in the 30 UTR of ART3 mRNA while one ARE class I motif was detected in the 3' UTR of ART4 mRNA.
引用
收藏
页码:53 / 57
页数:5
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