Effect of tandem repeated AUG codons on translation efficiency of eukaryotic mRNA carrying a short leader sequence |
| |
| Authors: | Motoaki Wakiyama Ichiro Hirao Izumi Kumagai and Kin-ichiro Miura |
| |
| Institution: | (1) Department of Industrial Chemistry, Faculty of Engineering, The University of Tokyo, 113 Hongo, Tokyo, Japan;(2) Present address: Institute for Biomolecular Science, Gakushuin University, 171 Mejiro, Tokyo, Japan;(3) Present address: Laboratory of Pharmaceutical Chemistry, Tokyo College of Pharmacy, 192-03 Hachioji, Tokyo, Japan |
| |
| Abstract: | The effect on translation of multiple copies of the initiation codon AUG at the initiation site in a eukaryotic mRNA carrying a short leader sequence was tested in translation experiments in vitro. DNA, corresponding to a chimeric mRNA sequence consisting of the 5 leader region of brome mosaic virus (BMV) RNA4 and the goat pre- -lactalbumin mRNA sequence, was prepared and transcribed in vitro using SP6 RNA polymerase. Site-directed mutagenesis was carried out to change the sequence around the initiation codon AUG. In a wheat germ translation system, the yield of protein obtained using the mRNA with a duplication of the AUG codons at the initiation site was 1.6 times that achieved when only one AUG was present. The rate of formation of the 80S initiation complex was measured by the ribosome binding assay using cycloheximide. A good correlation was observed between the ability to form the complex and translation efficiency. |
| |
| Keywords: | Eukaryotic mRNA Initiation codon AUG In vitro translation Translation efficiency |
| 本文献已被 SpringerLink 等数据库收录！ |
|
