Fig 1.
Growth of Symbiodinium CCMP2467 in f/2 liquid medium under antibiotic selection.
Results of Symbiodinium growth were from three biological replicates, each measured four times using a FlowCAM. The error bars indicate standard error of the mean (S16 File).
Fig 2.
S. cerevisiae growth under chloramphenicol selection.
“Parental” refers to the parental, untransformed S. cerevisiae yeast strain 31019b. “Empty vector” refers to the parental yeast strain transformed with a pYES2.1 vector carrying an eGFP coding region. “Chlo Resist” refers to the parental yeast strain transformed with a pYES2.1 vector carrying the chloramphenicol acyltransferase (CAT) resistance gene (ChloR). “+Chlo” indicates treatment of cultures with chloramphenicol. “+Ura” and “-URA” indicate the presence and absence of uracil supplement in the growth media respectively. Results are from five biological replicates. Asterisks indicate significantly different means with a p-value of < 0.01 using Student’s t-test with unequal variance. T-test was carried out with a significance value of 0.05. Error bars indicate standard error of the mean (S17 File).
Fig 3.
5’ RACE results of actin transcript to identify 5’ UTR sequence.
Fig 4.
Unique promoter architecture in Symbiodinium.
(A) A schematic view of the proposed promoter (TTTT-box) relative to the putative transcription start site, splice branch point and acceptor site, upstream of the coding region (CDS) in the genomic sequence of S. kawagutii genes. For comparison, premature mRNA and mature mRNA are also shown. Figure was adapted from Lin et al. (2015). (B) Genomic sequence of S. microadriaticum pAct promoter 175 bp upstream of the putative start codon, with promoter elements as described in (A).
Fig 5.
Vector maps for selected transformation plasmids.
A) Vector map for plasmid pAct-ChloR-ActT, constructed using the pCR2.1 vector backbone. B) Vector map for plasmid p35S-ChloR-NosT, constructed using the pCR4 vector backbone.
Table 1.
List of constructs.
Table 2.
Summary list of the number of experiments carried out grouped by transformation method.
Table 3.
List of constructs used for CS-153 transformation using silicon carbide whiskers.
Table 4.
List of primers used to construct expression cassettes for Symbiodinium expression vectors.