Development of a new cucumber mosaic virus-based plant expression vector with truncated 3a movement protein.
We have developed a Cucumber mosaic virus (CMV)-based expression vector for the production of heterologous proteins in plants. Cell-to-cell movement of CMV is dependent on the presence of coat protein (CP). Previous studies have shown that deletion of 33 amino acids (aa) from the carboxy-terminus of the 3a movement protein facilitates cell-to-cell movement that is independent of CP. The CMV-based expression vector that we have designed utilizes this truncated 3a protein, allowing the expression of target genes from the strong CP subgenomic promoter and without the need for providing CP in trans for cell-to-cell spread. Using this vector we achieved expression levels of ~450 mg/kg leaf tissue of green fluorescent protein (GFP) when the vector was delivered into Nicotiana benthamiana plants by agroinfiltration. Human growth hormone (hGH), on the other hand, accumulated to ~170 mg/kg of leaf tissue when the same approach was used to deliver the vector.
Fujiki
Masaaki
M
Fraunhofer USA Center for Molecular Biotechnology, 9 Innovation Way, Suite 200, Newark, DE 19711, USA.
Kaczmarczyk
Jon F
JF
Yusibov
Vidadi
V
Rabindran
Shailaja
S
D017799:Cucumovirus / Q000235:genetics* / Q000502:physiology*; D015967:Gene Expression Regulation, Viral; D005818:Genetic Engineering; D005822:Genetic Vectors / Q000235:genetics*; D049452:Green Fluorescent Proteins / Q000378:metabolism; D019382:Human Growth Hormone / Q000235:genetics; D006801:Humans; D053830:Plant Viral Movement Proteins / Q000235:genetics / Q000378:metabolism*; D011994:Recombinant Proteins / Q000378:metabolism; D017384:Sequence Deletion; D014026:Nicotiana / Q000821:virology
D053830:Plant Viral Movement Proteins; D011994:Recombinant Proteins; D019382:Human Growth Hormone; D049452:Green Fluorescent Proteins