Abstract
Protoplasts isolated from pea leaves (Pisum sativum L. cv. Hurst Greenshaft) were electroporated in the presence of plasmid pDR#1, which contains the rat liver ATP:citrate lyase gene fused to a duplex 35S cauliflower mosaic virus promoter with a transit peptide sequence of the Rubisco small subunit. The level of enzyme expression and viability of protoplasts were both influenced by polyethylene glycol treatment before electroporation. Under the optimised electroporation conditions, an average increase of ATP:citrate lyase activity of 14% was observed in the transfected cells after 24 h, with a similar magnitude of change in the abundance of the corresponding mRNA. Immunoblot analysis confirmed the correct expression and targeting of ATP:citrate lyase protein in the chloroplasts of pea protoplasts. These results provide a basis for the establishment of a procedure for targeting heterologous protein into pea plastids in the presence of a transit peptide.
Similar content being viewed by others
Author information
Authors and Affiliations
Additional information
Received: 14 June 1996 / Revision received: 24 November 1996 / Accepted: 4 January 1997
Rights and permissions
About this article
Cite this article
Rangasamy, D., Ratledge, C. & Woolston, C. Plastid targeting and transient expression of rat liver ATP: citrate lyase in pea protoplasts. Plant Cell Reports 16, 700–704 (1997). https://doi.org/10.1007/s002990050305
Issue Date:
DOI: https://doi.org/10.1007/s002990050305