Introduction

Paromomycin belongs to a group of aminoglycoside antibiotics such as neomycin or dibekacin. These aminoglycosides are capable of inhibiting the eukaryotic translation, by binding within the large and small subunit of the 80S ribosome. This property allows paromomycin to be used as selection marker for C.reinhardtii. For the selection process to work, one must consider a way to implement a paromomycin resistance in Chlamydomonas. The bacteria Stretpomyces rimous carries the aminoglycoside 3’-phosphotransferase encoded in the so called aphVIII gene. This enzyme catalyses the transfer of the gamma-phosphate of ATP to the hydroxyl group in 3’ position of the paromomycin molecule and allows the carrier of the gene to develop a resistance to paromomycin (Sizova et al. 2001). We used this resistance as a screening method for most of our transformations. During our research we discovered, that this resistance gene was already contributed to the iGem Registry. We wanted to improve this part by changing the codon usage and see if this improvement would end up in a higher expression of the aminoglycoside 3’-phosphotransferase and therefore in a better resistance to paramomycin.

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Modified Codon Usage of Paromomycin Plasmid

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