Transfection of the protozoan parasite Perkinsus marinus

2008 ◽  
Vol 157 (1) ◽  
pp. 44-53 ◽  
Author(s):  
José A. Fernández-Robledo ◽  
Zhuoer Lin ◽  
Gerardo R. Vasta
Keyword(s):  
Protozoan Parasite ◽  
Perkinsus Marinus

Host oyster tissue extracts modulate in vitro protease expression and cellular differentiation in the protozoan parasite, Perkinsus marinus

Parasitology ◽  
2003 ◽  
Vol 126 (4) ◽  
pp. 293-302 ◽  
Author(s):  
E. A. MACINTYRE ◽  
C. G. EARNHART ◽  
S. L. KAATTARI
Keyword(s):  
Serine Proteases ◽  
Protozoan Parasite ◽  
Eastern Oyster ◽  
Defined Medium ◽  
Tissue Homogenate ◽  
Oyster Tissue ◽  
Perkinsus Marinus ◽  
Tissue Extracts

Perkinsus marinus is responsible for a chronic disease (Dermo) of the Eastern oyster, Crassostrea virginica. In order to simulate the in vivo environment more closely, a chemically defined medium (JL-ODRP-3) was supplemented with tissue homogenate extracts or plasma from oysters possessing varying degrees of susceptibility to P. marinus infection. In media supplemented with extracts from highly susceptible oysters (C. virginica), P. marinus cells secreted elevated amounts of a set of low molecular weight serine proteases (LMP: 30–45 kDa) as assessed by enhanced digestion within gelatin-substrate SDS–PAGE gels. Oyster species of low susceptibility (C. gigas and C. ariakensis) did not exhibit this ability to upregulate P. marinus LMP expression. Oyster extract supplementation also led to pronounced changes in P. marinus cellular morphology, such that the cells were comparable to those observed within naturally infected oysters.

scholarly journals CRISPR/Cas9 Ribonucleoprotein-Based Genome Editing Methodology in the Marine Protozoan Parasite Perkinsus marinus

2021 ◽  
Vol 9 ◽  
Author(s):  
Raghavendra Yadavalli ◽  
Kousuke Umeda ◽  
Hannah A. Waugh ◽  
Adrienne N. Tracy ◽  
Asha V. Sidhu ◽  
...  
Keyword(s):  
Heterologous Gene Expression ◽  
Protozoan Parasite ◽  
Close Relative ◽  
Perkinsus Marinus ◽  
Post Transfection ◽  
Expression Platform ◽  
C Terminus ◽  
Gene Expression Platform ◽  
Optimized Protocol ◽  
Dermo Disease

Perkinsus marinus (Perkinsozoa), a close relative of apicomplexans, is an osmotrophic facultative intracellular marine protozoan parasite responsible for “Dermo” disease in oysters and clams. Although there is no clinical evidence of this parasite infecting humans, HLA-DR40 transgenic mice studies strongly suggest the parasite as a natural adjuvant in oral vaccines. P. marinus is being developed as a heterologous gene expression platform for pathogens of medical and veterinary relevance and a novel platform for delivering vaccines. We previously reported the transient expression of two rodent malaria genes Plasmodium berghei HAP2 and MSP8. In this study, we optimized the original electroporation-based protocol to establish a stable heterologous expression method. Using 20 μg of pPmMOE[MOE1]:GFP and 25.0 × 106P. marinus cells resulted in 98% GFP-positive cells. Furthermore, using the optimized protocol, we report for the first time the successful knock-in of GFP at the C-terminus of the PmMOE1 using ribonucleoprotein (RNP)-based CRISPR/Cas9 gene editing methodology. The GFP was expressed 18 h post-transfection, and expression was observed for 8 months post-transfection, making it a robust and stable knock-in system.

scholarly journals Transient Expression of Plasmodium berghei MSP8 and HAP2 in the Marine Protozoan Parasite Perkinsus marinus

10.1645/16-88 ◽  
2017 ◽  
Vol 103 (1) ◽  
pp. 118-122 ◽  
Author(s):  
Emma R. Cold ◽  
Gerardo R. Vasta ◽  
José A. Fernández Robledo
Keyword(s):  
Transient Expression ◽  
Plasmodium Berghei ◽  
Protozoan Parasite ◽  
Perkinsus Marinus
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