Abstract
Giardia lamblia is an important causative agent of persistent diarrhea in humans, domestic animals, and cattle. Basic research is usually performed with the strain WBC6 and includes genetic manipulations such as transfections. Here, we investigate how transfection with a plasmid causing stable expression of a foreign gene affects the whole proteome pattern. Using shotgun mass spectrometry, we compare the proteomes of untransfected trophozoites to trophozoites transfected with Escherichia coli glucuronidase A (GusA). Besides GusA, which is detected in the transfected trophozoites only, the proteomes of untransfected and transfected trophozoites differ by 132 differentially expressed proteins. In particular, transfection induces antigenic variation. Since transfection causing stable expression affects the proteome pattern, transfection experiments should take into account this effect. Due to a unique peptide panel, GusA is an example for a suitable internal standard for experiments involving transfected cells. Data are available via ProteomeXchange with identifier PXD022565.
Highlights
Giardiasis can be regarded as a zoonosis since it occurs in humans as well as in other mammals (Thompson, 2004)
Shotgun mass spectrometry of the proteomes of untransfected G. lamblia WBC6 trophozoites (WT) and of trophozoites transfected with a plasmid containing the S. alboniger puromycin-N-acetlytransferase (Pac) as a resistance marker and the E. coli glucuronidase A gene (Gus A) resulted in the identification of 20’091 unique peptides matching to 1’705 proteins
12 unique peptides matching to glucuronidase A (GusA) were consistently detected in the transfected trophozoites, but not in the WT
Summary
G. duodenalis, G. intestinalis), is an early diverging, anaerobic eukaryote (Müller and Müller, 2016; Cernikova et al, 2018) causing persistent diarrhea, especially in regions with low hygienic standards (Hemphill et al, 2019). After stomach passage, ingested cysts transform into trophozoites colonizing the duodenum. Hosts in good physical condition face the strongest symptoms of giardiasis 1 week post infection, and recover within two to three weeks. The infection becomes chronical causing severe damage of the intestinal epithelium, which may result in the development of irritable bowel syndrome (Allain et al, 2017; Litleskare et al, 2018). Giardiasis can be regarded as a zoonosis since it occurs in humans as well as in other mammals (Thompson, 2004)
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