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. Author manuscript; available in PMC: 2013 Nov 14.
Published in final edited form as: Mol Biochem Parasitol. 2008 Jun 7;161(1):10.1016/j.molbiopara.2008.05.006. doi: 10.1016/j.molbiopara.2008.05.006

Table 1.

Single-locus targeting (pRPa) and other complementary constructs.

Constructs Namea Selectable
Markerb 		Expression
Cassette		 Tet-
regulated		 Linearise
with
pRPa pRPaiSL HYG Stem-loop RNAic Yes AscI
pRPaiGx HYG GFPx Yes AscI
pRPaixG HYG xGFP Yes AscI
pRPaiMx HYG Mycx Yes AscI
pRPaix6Mx HYG x6Mycx Yes AscI
pRP d pRPiMx BSD Mycx Yes NotI
pRPix6Mx BSD x6Mycx Yes NotI
pNATe pNATxG BSD xGFP No f
pNATx12M BSD x12Myc No f
pNATGx BSD GFPx No f
pNAT6Mx BSD 6Mycx No f
a

RP, RRNA promoter controls the expression cassette; NAT, native locus controls expression; a, denotes AscI sites; i, inducible; SL, stem-loop; x, selected ORF; G, eGFP; M, cMyc epitope.

b

HYG, hygromycin-B phosphotransferase; BSD, blasticidin-S deaminase.

c

Different multiple cloning sites available: MCS1/2, MCS3/4 (see Fig. 1).

d

pRP constructs can integrate at any RRNA spacer locus and yield variable levels of expression upon induction in different clones.

e

pNAT allows expression of a tagged ORF from it’s native locus; see [2-4].

f

The restriction enzyme used depends upon the specific ORF targeted.