Skip to main page content
Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
, 153 (9), 1667-75

Insertion in the Coding Region of the Movement Protein Improves Stability of the Plasmid Encoding a Tomato Mosaic Virus-Based Expression Vector

Affiliations

Insertion in the Coding Region of the Movement Protein Improves Stability of the Plasmid Encoding a Tomato Mosaic Virus-Based Expression Vector

Koji Dohi et al. Arch Virol.

Abstract

A major obstacle in the genetic manipulation of tomato mosaic virus (ToMV) is the instability of the plasmid containing the infectious full-length cDNA of the ToMV vector, which often prevents the subcloning of a foreign gene of interest into the vector. We found that an insertion of a 0.3-1.6-kbp DNA fragment in the movement protein (MP) coding region effectively attenuated bacterial toxicity of the plasmid and greatly increased plasmid yield. Accumulation of a modified ToMV containing a 0.3-kb insertion in the MP coding region was comparable to that of a modified ToMV without an insertion in tobacco BY-2 protoplasts, while an insertion more than 0.6 kb significantly reduced accumulation of the viral RNA. The modified ToMV vector containing a 0.3-kb insertion was easily manipulated to introduce a coding sequence for human interferon-gamma (HuIFN-gamma) and successfully utilized to produce HuIFN-gamma in both BY-2 protoplasts and transgenic BY-2 cells.

Similar articles

See all similar articles

Cited by 5 PubMed Central articles

Publication types

Substances

LinkOut - more resources

Feedback