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Generation and Analysis of K173cl1-GFP-Luc Constructs in Schizont Stage Plasmodium

This study describes the generation of two distinct GFP-Luciferase integration constructs, K173cl1-GFP-Lucschiz-a and K173cl1-GFP-Lucschiz-b, in Plasmodium species via single and double cross-over strategies. These constructs were targeted to the c/d-rRNA locus on chromosomes 5 and 6. Southern blot analysis was performed to confirm successful integration using an ANKA 3'-UTR dhfr/ts probe. The results demonstrate the efficacy of both integration methods, providing valuable tools for further studies in Plasmodium biology and gene expression monitoring.

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Generation and Analysis of K173cl1-GFP-Luc Constructs in Schizont Stage Plasmodium

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  1. 4x106 1,2x106 Supp Figure 1 Generation of K173cl1-GFP-Lucschiz-a (gfp-luc integration by single cross-over) A gfp-luc K construct pL0028 SM 3'ssu pUC ama1 3’dhfr 5'ssu c/d-rRNA locus (chromsome 5/6) c/d -ets ssu lsu 5.8S gfp-luc c/d-rRNA locus after integration SM c/d -ets ssu’ ama1 3’dhfr ssu B Generation of K173cl1-GFP-Lucschiz-b (gfp-luc integration by double cross-over) gfp-luc S S construct pL1403 3'dhfr ama1 5‘ssu 3’ssu c/d-rRNA locus (chromsome 5/6) c/d -ets ssu lsu 5.8S Δssu Δssu gfp-luc c/d-rRNA locus after integration c/d -ets ama1 3’dhfr Southern analysis of separated chromosomes (Probe: ANKA_3’-UTR dhfr/ts) C K173cl1 -GFP-Lucschiz-b K173cl1 -GFP-Lucschiz-a ANKAwt ANKAwt K173cl1 - chr 7 - chr 7 chr 5/6 - chr 5/6 - chr 7 - chr 7 - D K173cl1 GFP-Lucschiz-b K173cl1 GFP-Lucschiz-a

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