CC-5168 cw15 ∆psbH [strain TN72]
$30.00
From Saul Purton, University College London, February 2016
This mutant is a chloroplast transformant in which the aadA (SpcR) cassette has been inserted between the BstXI site in the middle of the psbH coding sequence and the MluI site in the psbH-trnE2 intergenic region, thereby deleting a 0.36 kb genomic region including a large part of psbH. The aadA gene is in the same orientation as psbH.
Transformant is PSII-minus (i.e. acetate-requiring) due to the loss of PsbH, and spectinomycin resistant due to the presence of AadA. TN72 serves as a recipient strain for chloroplast transformation using the pASapI, pSRSapI and pWUCA2 expression vectors developed in the Purton lab.
Young RE and Purton S (2014) Cytosine deaminase as a negative selectable marker for the microalgal chloroplast: a strategy for the isolation of nuclear mutations that affect chloroplast gene expression. Plant J 80:915–925
Wannathong et al. (2016). New tools for chloroplast genetic engineering allow the synthesis of human growth hormone in the green alga Chlamydomonas reinhardtii. Applied Microbiology and Biotechnology. In press.
Young REB, Purton S (2016) Codon reassignment to facilitate genetic engineering and biocontainment in the chloroplast of Chlamydomonas reinhardtii. Plant Biotechnol J [Epub ahead of print]. doi: 10.1111/pbi.12490