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. Author manuscript; available in PMC: 2013 Nov 9.
Published in final edited form as: Methods Cell Biol. 2009 Dec 4;93:10.1016/S0091-679X(08)93002-8. doi: 10.1016/S0091-679X(08)93002-8

Fig. 4. Schematic Illustration outlining procedure for C-terminal in situ tagging of T. brucei genes via “knock-in” approach using the pMOTag vector series (Oberholzer et al. 2006).

Fig. 4

The tagging-integration construct is generated using PCR with chimeric primers including target gnee ORF (blue) or 3′UTR (gray) at the 5′ end and pMOTag sequences at the three prime end. Alternitively, the integration construct is generated by cloning ORF and UTR sequences into the pMOTag multicloning sites (MCS). The tagging-integration construct is integrated at the endogenous chromosomal locus by homologous recombination. Analogous pMOTag vectors are used for tagging at theN-terminus (Supplemental Fig. 1).