Analysis of the 3' UTR of the ART3 and ART4 gene by 3' inverse RACE-PCR.
3' Rapid amplification of cDNA ends (3' RACE) is a polymerase chain reaction (PCR) based technique which has been developed to analyse 3' 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 3' 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 3' UTR of ART3 mRNA while one ARE class I motif was detected in the 3' UTR of ART4 mRNA.
Friedrich
Maik
M
Institute of Biologie II, Department of Immunobiology, University of Leipzig, Germany.
Grahnert
Andreas
A
Hauschildt
Sunna
S
D020413:3' Untranslated Regions; D036002:ADP Ribose Transferases / Q000235:genetics*; D001483:Base Sequence; D002460:Cell Line; D017931:DNA Primers / Q000235:genetics; D058851:GPI-Linked Proteins; D006801:Humans; D008297:Male; D008565:Membrane Proteins / Q000235:genetics*; D008969:Molecular Sequence Data; D009206:Myocardium / Q000201:enzymology; D016133:Polymerase Chain Reaction / Q000379:methods*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D013737:Testis / Q000201:enzymology
D020413:3' Untranslated Regions; D017931:DNA Primers; D058851:GPI-Linked Proteins; D008565:Membrane Proteins; D012333:RNA, Messenger; D036002:ADP Ribose Transferases; C105233:ART3 protein, human; C105234:ART4 protein, human