E. Kobatake et al., Stabilization and translation of immobilized mRNA on latex beads for cell-free protein synthesis system, APPL BIOC B, 76(3), 1999, pp. 217-227
The stability of immobilized mRNA against ribonucleases was investigated in
a cell-free protein synthesis system. The plasmid-encoding protein A with
the 20-mer poly(A) tail under the control of T7 promoter was constructed, a
nd the corresponding mRNA was synthesized by T7 RNA polymerase reaction. Th
e resulting mRNA was immobilized on oligo(dT)-immobilized latex beads by hy
bridization utilizing the poly(A) tail of mRNA at the 3'-terminus. The mRNA
was stabilized against three types of nucleases (3'-OH exonuclease, 5'-OH
exonuclease, and endonuclease) by immobilization. Translation of immobilize
d mRNA with a continuous-flow cell-free protein-synthesizing system from Sa
ccharomyces cerevisiae was ascertained. Reusability of the immobilized mRNA
as genetic information was also examined.