Transient recombinant expression of highly immunogenic CagA, VacA and NapA in Nicotiana benthamiana

Abstract

Highlights A platform for rapid expression of genes through agro-infiltration is a suitable alternative to the laborious process of generating stable transgenic lines for producing recombinant immunogenic antigens in plants. This study has successfully developed and demonstrated a reproducible transformation protocol for rapid expression of Helicobacter pylori recombinant CagA, VacA and NapA antigens in Nicotiana benthamiana . This is the first study of recombinant expression CagA, VacA and NapA gene of Helicobacter pylori in Nicotiana benthamiana via syringe-assisted A grobacterium infiltration. A syringe infiltration of a four to five weeks old Nicotiana benthamiana plant with Agrobacterium tumefaciens subtype EHA105 was optimal to produce the maximum mRNA levels of CagA, VacA and NapA mRNA in leaf at the third-day post-Agro-infiltration.

Stable expression ofHelicobacter pylori cagAoncoprotein in brinjal

“…The cagA transgenic brinjal leaves extracts were diluted as 1:5 and 1:10 in sodium bicarbonate (pH 9.8) and coated to the wells of the ELISA plate by adding 4.5-5 mg/mL concentration of antigen solution and incubating at 37 °C for 2 h, followed by three washes with washing buffer (0.1 percent Tween 20 in PBS) for 5 min each time (Barzigar et al, 2022). The plate was blocked for 90 minutes with 2 percent skim milk, then cleaned three times with washing solution.…”