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Applied Biochemistry and Biotechnology

, Volume 76, Issue 3, pp 217–227 | Cite as

Stabilization and translation of immobilized mRNA on latex beads for cell-free protein synthesis system

  • Eiry Kobatake
  • Akira Ebisawa
  • Orie Asaka
  • Yasuko Yanagida
  • Yoshihito Ikariyama
  • Masuo AizawaEmail author
Article

Abstract

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, and the corresponding mRNA was synthesized by T7 RNA polymerase reaction. The resulting mRNA was immobilized on oligo(dT)-immobilized latex beads by hybridization 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 immobilized mRNA with a continuous-flow cell-free protein-synthesizing system from Saccharomyces cerevisiae was ascertained. Reusability of the immobilized mRNA as genetic information was also examined.

Index Entries

Stabilization of mRNA immobilized mRNA cell-free protein synthesis continuous flow system Saccharomyces cerevisiae protein A 

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Copyright information

© Humana Press Inc 1999

Authors and Affiliations

  • Eiry Kobatake
    • 1
  • Akira Ebisawa
    • 1
  • Orie Asaka
    • 1
  • Yasuko Yanagida
    • 1
  • Yoshihito Ikariyama
    • 2
  • Masuo Aizawa
    • 1
    Email author
  1. 1.Department of Bioengineering, Faculty of Bioscience and BiotechnologyTokyo Institute of TechnologyYokohamaJapan
  2. 2.Research InstituteNational Rehabilitation CenterTokorozawaJapan

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