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. 2016 Oct 20;6(12):4115–4125. doi: 10.1534/g3.116.033035

Table 4. Plasmids available for transgene expression using the one-promoter system.

Plasmida Promoterb Upstream Genec Selectable Markerd
pMO449e HSP70A/RBCS2 CrVENUS-3FLAG APHVIII
pMO470e HSP70A/RBCS2 CrVENUS-3FLAG APHVIII
pMO471e HSP70A/RBCS2 CrVENUS-3FLAG APHVIII
pMO488 HSP70A/RBCS2 CrVENUS APHVIII
pMO490 HSP70A/RBCS2 sfGFP-3FLAG APHVIII
pMO519 HSP70A/RBCS2 CrmCHERRY APHVIII
pMO520 HSP70A/RBCS2 CrmCHERRY-3FLAG APHVIII
pMO507 PSAD CrVENUS-3FLAG APHVIII
pMO508 TUB2 CrVENUS-3FLAG APHVIII
pMO561f TUB2 CrVENUS-3FLAG APHVIII
a

Available plasmids with one-promoter expression constructs of the general structure shown in Figure 1B.

b

Except as indicated in note f, all promoters are followed by the first three codons and intron of RBCS2.

c

All genes indicated have been codon optimized for Chlamydomonas. These genes can be replaced in toto by HpaI–StuI fragments containing other GOIs whose expression/overexpression is desired. Fluorescence tagging of a gene product of interest can be achieved by inserting the coding sequence, in frame, at the HpaI site (C-terminal tagging) or the StuI site (N-terminal tagging).

d

Other selectable markers could be inserted as NdeI–BamHI fragments. To date, however, we have not had success with selectable markers other than APHVIII (see text).

e

These plasmids differ only in the short linker between CrVENUS-3FLAG and APHVIII (see Figure 2D and Table 2).

f

Same as pMO508 except that the RBCS2 ATG start codon has been changed to TTG. This allowed expression of CrVENUS-3FLAG, presumably from its own start codon (i.e., without the additional amino acids from RBCS2).