Stabilization of mRNA in the cell-free translation system from Escherichia coli.
In the RNA directed cell-free protein synthesizing system from E. coli, there is a problem of contaminating 3'-exonucleases which attack the mRNAs. Thus, we tried the following two methods to stabilize mRNAs against nucleases: (A) To use mRNAs having hairpin structures at their termini and (B) To hybridize mRNAs with small DNA fragments to the 3'-termini of mRNAs. It was found that degradation of a mRNA was inhibited by the method B rather than the method A in the translation system.
Hirao
I
I
Department of Industrial Chemistry, Faculty of Engineering, University of Tokyo, Japan.
Yoshizawa
S
S
Miura
K
K
D000595:Amino Acid Sequence; D001483:Base Sequence; D002474:Cell-Free System; D004269:DNA, Bacterial / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics*; D005092:Exonucleases / Q000378:metabolism; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D009693:Nucleic Acid Hybridization; D009842:Oligopeptides / Q000235:genetics; D010675:Pheromones / Q000235:genetics; D014176:Protein Biosynthesis; D012329:RNA, Bacterial / Q000378:metabolism*; D012333:RNA, Messenger / Q000378:metabolism*
D004269:DNA, Bacterial; D009842:Oligopeptides; D010675:Pheromones; D012329:RNA, Bacterial; D012333:RNA, Messenger; C043691:cAD1 bacterial sex hormone; D005092:Exonucleases