scholarly journals AI239 and RB95 antibodies recognize a GST-tagged recombinant protein by immunofluorescence

2020 ◽  
Vol 3 (2) ◽  
pp. e132
Author(s):  
Wanessa C Lima ◽  
Pierre Cosson
Keyword(s):  
Recombinant Protein ◽  
Hela Cells ◽  
Glutathione S Transferase

AI239 and RB95 antibodies against the Glutathione S-transferase (GST) protein recognize a GST-tagged human TAC protein by immunofluorescence in paraformaldehyde-fixed HeLa cells; AF209, AF212 and RB94 do not.

scholarly journals Inhibition of Fungal Appressorium Formation by Pepper (Capsicum annuum) Esterase

2001 ◽  
Vol 14 (1) ◽  
pp. 80-85 ◽  
Author(s):  
Young Soon Kim ◽  
Hyun Hwa Lee ◽  
Moon Kyung Ko ◽  
Chae Eun Song ◽  
Cheol-Yong Bae ◽  
...  
Keyword(s):  
Recombinant Protein ◽  
Capsicum Annuum ◽  
Magnaporthe Grisea ◽  
Dependent Manner ◽  
Appressorium Formation ◽  
Glutathione S Transferase ◽  
Porcine Liver ◽  
Esterase Gene ◽  
Dose Dependent ◽  
Dependent Signaling Pathway

A pepper esterase gene (PepEST) that is highly expressed during an incompatible interaction between pepper (Capsicum annuum) and the anthracnose fungus Colletotrichum gloeosporioides has been previously cloned. Glutathione-S-transferase-tagged recombinant PepEST protein expressed in Escherichia coli showed substrate specificity for p-nitrophenyl esters. Inoculation of compatible unripe pepper fruits with C. gloeosporioides spores amended with the recombinant protein did not cause anthracnose symptoms on the fruit. The recombinant protein has no fungicidal activity, but it significantly inhibits appressorium formation of the anthracnose fungus in a dose-dependent manner. An esterase from porcine liver also inhibited appressorium formation, and the recombinant protein inhibited appressorium formation in the rice blast fungus, Magnaporthe grisea. Inhibition of appressorium formation in M. grisea by the recombinant protein was reversible by treatment with cyclic AMP (cAMP) or 1,16-hexadecanediol. The results suggest that the recombinant protein regulates appressorium formation by modulating the cAMP-dependent signaling pathway in this fungus. Taken together, the PepEST esterase activity can inhibit appressorium formation of C. gloeosporioides, which may result in protection of the unripe fruit against the fungus.

scholarly journals The AI196 antibody recognizes a SPOT-tagged recombinant protein by immunofluorescence

2020 ◽  
Vol 3 (2) ◽  
pp. e136
Author(s):  
Wanessa C Lima ◽  
Pierre Cosson
Keyword(s):  
Recombinant Protein ◽  
Hela Cells ◽  
Spot Tag

The AI196 antibody against the SPOT tag recognizes a SPOT-tagged human TAC protein by immunofluorescence in paraformaldehyde-fixed HeLa cells.

scholarly journals AE391 and AF291 antibodies recognize an HA-tagged recombinant protein by immunofluorescence

2020 ◽  
Vol 3 (2) ◽  
pp. e133
Author(s):  
Wanessa C Lima ◽  
Pierre Cosson
Keyword(s):  
Recombinant Protein ◽  
Hela Cells

AE391 and AF291 antibodies against the HA tag recognize an HA-tagged human TAC protein by immunofluorescence in paraformaldehyde-fixed HeLa cells.

scholarly journals The AI215 antibody recognizes an EPEA-tagged recombinant protein by immunofluorescence

2020 ◽  
Vol 3 (2) ◽  
pp. e130
Author(s):  
Wanessa C Lima ◽  
Pierre Cosson
Keyword(s):  
Recombinant Protein ◽  
Hela Cells

The AI215 antibody against the EPEA tag recognizes an EPEA-tagged human TAC protein by immunofluorescence in paraformaldehyde-fixed HeLa cells.

scholarly journals Plasmodium falciparum Phospholipase C Hydrolyzing Sphingomyelin and Lysocholinephospholipids Is a Possible Target for Malaria Chemotherapy

2001 ◽  
Vol 195 (1) ◽  
pp. 23-34 ◽  
Author(s):  
Kentaro Hanada ◽  
Nirianne Marie Q. Palacpac ◽  
Pamela A. Magistrado ◽  
Ken Kurokawa ◽  
Ganesh Rai ◽  
...  
Keyword(s):  
Plasmodium Falciparum ◽  
Recombinant Protein ◽  
Phospholipase C ◽  
Dependent Manner ◽  
Glutathione S Transferase ◽  
Trophozoite Stage ◽  
Severe Impairment ◽  
Active Transcription ◽  
Biochemical Analyses ◽  
Dose Dependent

Sphingomyelinase (SMase) is one of the principal enzymes in sphingomyelin (SM) metabolism. Here, we identified a Plasmodium falciparum gene (PfNSM) encoding a 46-kD protein, the amino acid sequence of which is ∼25% identical to that of bacteria SMases. Biochemical analyses of the recombinant protein GST-PfNSM, a fusion protein of the PfNSM product with glutathione-S-transferase, reveal that this enzyme retained similar characteristics in various aspects to SMase detected in P. falciparum–infected erythrocytes and isolated parasites. In addition, the recombinant protein retains hydrolyzing activity not only of SM but also of lysocholinephospholipids (LCPL) including lysophosphatidylcholine and lysoplatelet-activating factor, indicating that PfNSM encodes SM/LCPL-phospholipase C (PLC). Scyphostatin inhibited SM/LCPL-PLC activities of the PfNSM product as well as the intraerythrocytic proliferation of P. falciparum in a dose-dependent manner with ID50 values for SM/LCPL-PLC activities and the parasite growth at 3–5 μM and ∼7 μM, respectively. Morphological analysis demonstrated most severe impairment in the intraerythrocytic development with the addition of scyphostatin at trophozoite stage than at ring or schizont stages, suggesting its effect specifically on the stage progression from trophozoite to schizont, coinciding with the active transcription of PfNSM gene.

scholarly journals GST-TAT-SOD: Cell Permeable Bifunctional Antioxidant Enzyme—A Potential Selective Radioprotector

2016 ◽  
Vol 2016 ◽  
pp. 1-13 ◽  
Author(s):  
Jianru Pan ◽  
Huocong He ◽  
Ying Su ◽  
Guangjin Zheng ◽  
Junxin Wu ◽  
...  
Keyword(s):  
Recombinant Protein ◽  
Normal Liver ◽  
Liver Cells ◽  
Cell Penetrating Peptides ◽  
Radioprotective Effect ◽  
Glutathione S Transferase ◽  
Single Function ◽  
Cell Penetrating ◽  
Radioprotective Agent

Superoxide dismutase (SOD) fusion of TAT was proved to be radioprotective in our previous work. On that basis, a bifunctional recombinant protein which was the fusion of glutathione S-transferase (GST), SOD, and TAT was constructed and named GST-TAT-SOD. Herein we report the investigation of the cytotoxicity, cell-penetrating activity, and in vitro radioprotective effect of GST-TAT-SOD compared with wild SOD, single-function recombinant protein SOD-TAT, and amifostine. We demonstrated that wild SOD had little radioprotective effect on irradiated L-02 and Hep G2 cells while amifostine was protective to both cell lines. SOD-TAT or GST-TAT-SOD pretreatment 3 h prior to radiation protects irradiated normal liver cells against radiation damage by eliminating intracellular excrescent superoxide, reducing cellular MDA level, enhancing cellular antioxidant ability and colony formation ability, and reducing apoptosis rate. Compared with SOD-TAT, GST-TAT-SOD was proved to have better protective effect on irradiated normal liver cells and minimal effect on irradiated hepatoma cells. Besides, GST-TAT-SOD was safe for normal cells and effectively transduced into different organs in mice, including the brain. The characteristics of this protein suggest that it may be a potential radioprotective agent in cancer therapy better than amifostine. Fusion of two antioxidant enzymes and cell-penetrating peptides is potentially valuable in the development of radioprotective agent.

scholarly journals Generation and characterization of the VP1 recombinant protein of the chicken anemia virus

2019 ◽  
Vol 2019 (2) ◽  
pp. 12-20
Author(s):  
Михаил Грудинин ◽  
Mihail Grudinin ◽  
Андрей Комиссаров ◽  
Andrey Komissarov ◽  
Алина Гусейнова ◽  
...  
Keyword(s):  
Recombinant Protein ◽  
Recombinant Proteins ◽  
Polyclonal Antibodies ◽  
Chicken Anemia Virus ◽  
Glutathione S Transferase ◽  
Adenoviral Infection ◽  
Metal Affinity ◽  
Infectious Laryngotracheitis ◽  
Potential Component ◽  
Anemia Virus

The aim of this study is to obtain a VP1 recombinant protein of the chicken anemia virus, capable of specifically detecting antibodies in the blood sera of sick chickens. Materials and methods. Cloning of a fragment of the VP1 gene of an infectious anemia virus of chickens was performed in the expression plasmids pET15b and pGEX-3T in the context of reading the polyhistidine sequence and glutathione-S-transferase, respectively. The recombinant proteins 6HIS-ΔVP1 and GST-ΔVP1 expressed in E. coli Rosetta (DE3) strains were purified by metal affinity chromatography. Amino acid sequence of recombinant proteins was confirmed by mass spectrometry. The specificity of the interaction of recombinant proteins with polyclonal antibodies was determined by ELISA. The ability of the recombinant 6HIS-ΔVP1 protein to detect antibodies in field and blood sera of SPF chickens was evaluated by indirect ELISA. To control the specificity of the antigen, the immune sera of birds for viruses of infectious bronchitis, Newcastle disease, infectious laryngotracheitis, adenoviral infection, Mycoplasma gallisepticum, and negative serum of chickens were used. Results. The recombinant VP1 protein of chicken anemia virus containing a polyhistidine tag (6HIS-∆VP1) was obtained. It was shown that this recombinant protein is able to specifically detect antibodies in the blood sera of sick chickens. Conclusion. The obtained recombinant protein 6HIS-ΔVP1 can be used to detect antibodies to the chicken anemia virus in the serum of sick chickens, and can also be considered as a potential component of vaccines against this virus.

Expression of recombinant protein in cho and hela cells and its follow-up using EGF reporter gene

2006 ◽  
pp. 55-59
Author(s):  
V. Hendrick ◽  
D. Ribeiro Sousa ◽  
A. R. dos Santos Pedregal ◽  
C. Bassens ◽  
P. Rigaux ◽  
...  
Keyword(s):  
Recombinant Protein ◽  
Reporter Gene ◽  
Hela Cells
Sign in / Sign up

Export Citation Format

Share Document