Removal of the native uORF from the Pf86 plasmid

Chhaminder Kaur; Mayank Kumar; Swati Patankar

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Removal of the native uORF from the Pf86 plasmid

CK Chhaminder Kaur

MK Mayank Kumar

SP Swati Patankar

This method is extracted from research article: Parasitology, May 2020

Messenger RNAs with large numbers of upstream open reading frames are translated via leaky scanning and reinitiation in the asexual stages of Plasmodium falciparum

DOI: 10.1017/S0031182020000840

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Plasmid Pf86 was used to test the effect of synthetic uORFs on the expression of the firefly luciferase reporter gene. The native uORF present 474 bases upstream in the 5′ leader was removed by mutating the start codon (ATG) to TTG by SDM to get Pf86*, a plasmid devoid of any uORF in the 5′ leader. Primers used for this cloning are included in Additional File 1.

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