scholarly journals Development of polyclonal antisera against movement proteins from three poleroviruses infecting cucurbits

2020 ◽  
Vol 2 (1) ◽  
Author(s):  
Shao-Kang Zhang ◽  
Tian-Yu Zhao ◽  
Xing Shi ◽  
Yu-Zi Liu ◽  
Ying Wang ◽  
...  
Keyword(s):  
Movement Proteins ◽  
Polyclonal Antisera

scholarly journals Development of polyclonal antisera against movement proteins of the three poleroviruses infecting cucurbits and their serological relationships

2020 ◽  
Author(s):  
Shao-kang Zhang ◽  
Tian-yu Zhao ◽  
Xing Shi ◽  
Yu-zi Liu ◽  
Ying Wang ◽  
...  
Keyword(s):  
Prokaryotic Expression ◽  
Host Plants ◽  
Expression Vectors ◽  
Future Research ◽  
Serological Detection ◽  
Movement Proteins ◽  
Viral Movement ◽  
New Zealand White Rabbits ◽  
Polyclonal Antisera ◽  
Melon Aphid

Abstract Background: Cucurbit aphid-borne yellows virus (CABYV), Melon aphid-borne yellows virus (MABYV) and Suakwa aphid-borne yellows virus (SABYV) are three critical viruses infecting cucurbit crops. The preparation of specific antiserum against the virus is crucial for both the detection of virus and understanding the functions of the related genes. However, there is no report about detecting the three viruses using antisera against movement proteins (MP). Methods: In this study, we constructed prokaryotic expression vectors of the three viral movement proteins and transferred them into Escherichia coli strain Rosetta to purify the fusion proteins. Then the polyclonal antisera were obtained by immunizing New Zealand white rabbits. Western blotting was used to demonstrate the applicability of the three antisera. Results: We discovered that the titer of antiserum against MP CABYV reached to 1: 512000, and the titers of antisera against MP MABYV and MP SABYV reached to 1:256000. The optimized working concentration range for the three antisera was from 1:10000 to 1:64000. Both antisera against MP CABYV and MP MABYV could only react with the corresponding MP. The antiserum against MP SABYV not only had the strongest reaction with its MP but also could react with MP CABYV and MP MABYV at relative weaker levels and all the three antisera had no serological reactions with other poleroviruses tested. Furthermore, our results showed that the three antisera could specifically detect movement proteins both in Nicotiana benthamiana and cucumber leaves. Conclusions: We have established a sensitive system for detecting three poleroviruses infecting cucurbits by antisera against movement proteins, providing a material foundation for the future research on both the serological detection of viruses and the interaction mechanisms between the virus and host plants.

Use of monoclonal and polyclonal antibodies to detect prostatic acid phosphatase by immunoblotting.

1986 ◽  
Vol 32 (10) ◽  
pp. 1832-1835 ◽  
Author(s):  
P C Patel ◽  
L Aubin ◽  
J Côte
Keyword(s):  
Monoclonal Antibodies ◽  
Acid Phosphatase ◽  
Western Blot ◽  
Polyclonal Antibodies ◽  
Prostatic Acid Phosphatase ◽  
The Other ◽  
Dot Blot ◽  
Western Blots ◽  
Dot Blot Assay ◽  
Polyclonal Antisera

Abstract We investigated two techniques of immunoblotting--the Western blot and the dot blot--for use in detecting prostatic acid phosphatase (PAP, EC 3.1.3.2). We used polyclonal antisera to human PAP, produced in rabbits by hyperimmunization with purified PAP, and PAP-specific monoclonal antibodies in the immunoenzymatic protocols. We conclude that PAP can be readily detected by Western blots with use of polyclonal antisera, but not with monoclonal antibodies. On the other hand, using a dot blot assay, we could easily detect PAP with both polyclonal and monoclonal antibodies.

scholarly journals Evidence for Unknown Sarcocystis-Like Infection in Stranded Striped Dolphins (Stenella coeruleoalba) from the Ligurian Sea, Italy

Animals ◽  
2021 ◽  
Vol 11 (5) ◽  
pp. 1201
Author(s):  
Federica Giorda ◽  
Umberto Romani-Cremaschi ◽  
Antoinette E. Marsh ◽  
Carla Grattarola ◽  
Barbara Iulini ◽  
...  
Keyword(s):  
Neospora Caninum ◽  
Positive Staining ◽  
Ligurian Sea ◽  
Sarcocystis Neurona ◽  
Stenella Coeruleoalba ◽  
Polyclonal Antisera ◽  
Ligurian Coast ◽  
Sarcocystis Falcatula ◽  
The Brain ◽  
The Mediterranean Basin

Two striped dolphins (SD1, SD2), stranded along the Ligurian coast of Italy, were diagnosed with a nonsuppurative meningoencephalitis associated with previously undescribed protozoan tissue cysts. As tissue cysts were morphologically different from those of Toxoplasma gondii, additional histopathological, immunohistochemical, ultrastructural, and biomolecular investigations were performed, aiming to fully characterize the organism. Histopathology revealed the presence of large Sarcocystis-like tissue cysts, associated with limited inflammatory lesions in all CNS areas studied. IHC was inconclusive, as positive staining with polyclonal antisera did not preclude cross-reaction with other Sarcocystidae coccidia. Applied to each animal, 11 different PCR protocols precluded a neural infection by Sarcocystis neurona, Sarcocystis falcatula, Hammondia hammondi, and Neospora caninum. T. gondii coinfection was confirmed only in dolphin SD2. Sarcocystis sp. sequences, showing the highest homology to species infecting the Bovidae family, were amplified from SD1 myocardium and SD2 skeletal muscle. The present study represents the first report of Sarcocystis-like tissue cysts in the brain of stranded cetaceans along with the first description of Sarcocystis sp. infection in muscle tissue of dolphins from the Mediterranean basin.

scholarly journals Leaf Plasmodesmata Respond Differently to TMV, ToBRFV and TYLCV Infection

Plants ◽  
2021 ◽  
Vol 10 (7) ◽  
pp. 1442
Author(s):  
Yaarit Kutsher ◽  
Dalia Evenor ◽  
Eduard Belausov ◽  
Moshe Lapidot ◽  
Moshe Reuveni
Keyword(s):  
Cell Types ◽  
Plant Viruses ◽  
Tomato Leaf ◽  
Visual Localization ◽  
Tobacco Leaf ◽  
Signal Distribution ◽  
Movement Proteins ◽  
Significant Difference ◽  
Intracellular Mechanism ◽  
Leaf Curl Virus

Macromolecule and cytosolic signal distribution throughout the plant employs a unique cellular and intracellular mechanism called plasmodesmata (PD). Plant viruses spread throughout plants via PD using their movement proteins (MPs). Viral MPs induce changes in plasmodesmata’s structure and alter their ability to move macromolecule and cytosolic signals. The developmental distribution of a family member of proteins termed plasmodesmata located proteins number 5 (PDLP5) conjugated to GFP (PDLP5-GFP) is described here. The GFP enables the visual localization of PDLP5 in the cell via confocal microscopy. We observed that PDLP5-GFP protein is present in seed protein bodies and immediately after seed imbibition in the plasma membrane. The effect of three different plant viruses, the tobacco mosaic virus (TMV), tomato brown rugose fruit virus (ToBRFV, tobamoviruses), and tomato yellow leaf curl virus (TYLCV, begomoviruses), on PDLP5-GFP accumulation at the plasmodesmata was tested. In tobacco leaf, TMV and ToBRFV increased PDLP5-GFP amount at the plasmodesmata of cell types compared to control. However, there was no statistically significant difference in tomato leaf. On the other hand, TYLCV decreased PDLP5-GFP quantity in plasmodesmata in all tomato leaf cells compared to control, without any significant effect on plasmodesmata in tobacco leaf cells.

Production and use of monoclonal antibodies for identification of Bradyrhizobium japonicum strains

1988 ◽  
Vol 34 (1) ◽  
pp. 88-92 ◽  
Author(s):  
D. Velez ◽  
J. D. Macmillan ◽  
L. Miller
Keyword(s):  
Monoclonal Antibodies ◽  
Bradyrhizobium Japonicum ◽  
Chemical Nature ◽  
Periodate Oxidation ◽  
Enzyme Linked Immunosorbent Assay ◽  
Gram Negative Bacteria ◽  
Bacterial Cells ◽  
Gram Negative ◽  
Polyclonal Antisera ◽  
Somatic Antigens

Thirteen murine hybridomas capable of producing monoclonal antibodies to somatic antigens on Bradyrhizobium japonicum were developed and an indirect enzyme-linked immunosorbent assay was used to test reactivity of the antibodies against 20 strains of B. japonicum. Although polyclonal antisera from mice immunized with strains of B. japonicum reacted with bacterial cells of all 20 strains, individual monoclonals were more specific. Some antibodies reacted with as few as 2 and one with as many as 11 strains. On the basis of reactivity with the set of 13 monoclonal antibodies, the 20 strains of B. japonicum could be divided arbitrarily into five groups. Three of five monoclonal antibodies tested reacted with bacteroids taken directly from soybean nodules. One monoclonal bound to cells of five species of Rhizobium, but none of the 13 reacted with gram-negative bacteria representing six other genera. Treatment of cells with reagents and heat indicated the chemical nature of the antigens to five of the monoclonals. Antigen reactive with one antibody was destroyed by periodate oxidation indicating that it was a polysaccharide. Two antigens were probably proteins as they could be digested by trypsin and denatured by heat. Two others were inactivated by all three treatments suggesting they were glycoproteins.

Novel leukaemia markers

1995 ◽  
Vol 15 (6) ◽  
pp. 463-468
Author(s):  
Kingsley J. Micklem
Keyword(s):  
Monoclonal Antibodies ◽  
T Cell ◽  
Recombinant Protein ◽  
Follicular Lymphoma ◽  
B Lymphocytes ◽  
Synthetic Peptide ◽  
Paraffin Sections ◽  
Hematologic Disorders ◽  
Polyclonal Antisera ◽  
Chromosomal Defects

Using synthetic peptide or recombinant protein as immunising antigens we have produced monoclonal antibodies and polyclonal antisera directed against targets of particular interest in leukaemia diagnosis. In this way we have prepared reagents which recognise all T or all B lymphocytes in routinely fixed paraffin sections which are unique in this respect. We have also produced monoclonal antibodies to molecules potentially involved in specific neoplastic transformations, implicated by virtue of the involvement of their genes in chromosomal defects in these neoplasms. In particular, we have produced antibodies recognising bcl-2, involved in follicular lymphoma, tal-1, involved in T-cell acute leukaemias and HRX involved in a variety of hematologic disorders. The application of these reagents to diagnosis has so far proved useful. In addition their use outside the field of leukaemia diagnosis has proved to be even more important in some cases.

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