pPO3
$30.00
From Saul Purton, University College London, August 2023
Vector: pBluescript
Host strain: E. coli DH5α
Insert: The overall size of the plasmid is 9356 bp with the 2059 bp ptxD cassette flanked by homology arms of 2850 bp and 1523 bp (see plasmid map).
Selection: ampicillin resistant
A synthetic ptxD cassette was designed to serve as a selectable marker for chloroplast transformation, allowing selection on media containing phosphite as the P source. The cassette was optimised for expression in the chloroplast and cloned into plasmid pBa3-AX carrying a section of the chloroplast genome encompassing psaA-3, trnL and the 5’ region of ccsA. The ptxD cassette was inserted into a neutral intergenic locus using unique MluI and EcoRI sites between psaA-3 and trnL.
Changko S, Rajakumar PD, Young REB, Purton S. The phosphite oxidoreductase gene, ptxD as a bio-contained chloroplast marker and crop-protection tool for algal biotechnology using Chlamydomonas. Appl Microbiol Biotechnol. 2020 Jan;104(2):675-686. doi: 10.1007/s00253-019-10258-7. Epub 2019 Dec 2. PMID: 31788712; PMCID: PMC6943410.