Analysis of a series of λ cII::α1-antitrypsin (α1AT) gene fusions of different sizes showed that increased α1AT expression correlated with the stabilisation of a particular computer-predicted RNA secondary structure. Moreover, significant synthesis of unfused α1AT was achieved by reconstruction of this conformation to permit interaction between the upstream region of the ribosome-binding site and the first part of the α1 AT coding sequence. This high-level expression was dependent upon certain silent point mutations in the coding sequence, indicating that RNA primary and secondary structure determinants can operate in concert to dictate the efficiency of protein synthesis.
- recombinant DNA Gene fusion Point mutation λplasmid vector Gene cloning Emphysema
ASJC Scopus subject areas
- Structural Biology
- Molecular Biology
- Cell Biology