Stable transformation of unicellular green alga Coccomyxa subellipsoidea C-169 via electroporation

The effect of temperature conditions during growth on the transformation frequency of Coccomyxa subellipsoidea C-169 obtained by electroporation

“…Electrotransformation of C-169 cells was performed following an adaptation of a previously described procedure with some modifications [ 6 ], using a GenePulser apparatus (Bio-Rad Laboratories Limited, UK). Briefly, for transformation cells were washed with ice cold H 2 O three times and resuspended in ice cold H 2 O in final concentration of 1 × 5 × 10 8 cells mL −1 .…”

“…Recently, the utility of C. subellipsoidea C-169 as a potential biofuel producer has been intensively studied [ [3] , [4] , [5] ]. This prompted us to develop a system of transformation of C. subellipsoidea C-169 by electroporation [ 6 ]. Our previous research focused on finding the appropriate selection marker, determining the parameters of the electric field, and establishing the post-electroporation procedure for culturing the transformed cells.…”

Doing synthetic biology with photosynthetic microorganisms

“…Some members of the Chlorophyta are readily transformable. However, the members of Chlorophyta have thick cell walls and many strains need significant method optimization or cannot be transformed at all (Dehghani et al, 2018; Kania et al, 2020; Suttangkakul et al, 2019). Various methods are applicable for transformation of C. reinhardtii , including the glass bead method, particle bombardment, Agrobacterium ‐mediated conjugation, and electroporation being the main methods employed (Figure 2).…”

“…Electroporation is preferred, because of higher transformation efficiency, whereas particle bombardment is useful for transformation of mitochondria or chloroplast genomes (Wang et al, 2019). Similar methods have been used for the transformation of Chlorella (Yang et al, 2016), Coccomyxa (Kania et al, 2020; Kasai et al, 2018), and for Scenedesmus/Acutodesmus species (Dautor et al, 2014; Guo et al, 2013; Muñoz et al, 2018; Suttangkakul et al, 2019). In addition, an E. coli ‐mediated conjugation was reported for transformation of Acutodesmus obliquus (Muñoz et al, 2019).…”

“…Enhancer or enhancer-like elements, introns or retrotransposons have also been studied for improved expression (Yang et al, 2016). The endogenous RBCS promoter, and also the first intron of RBCS (Kania et al, 2020;Kasai et al, 2018) as well as the promoter of the elongation factor 1 alpha gene have been used to drive gene expression in Coccomyxa species (Kasai et al, 2017). A marker recycling system and marker-free engineering have also been reported (Kasai et al, 2017(Kasai et al, , 2018.…”