Molecular and recombinant characterization of major surface protein 5 from Anaplasma marginale

Watthanadirek, Amaya; Junsiri, Witchuta; Minsakorn, Sutthida; Poolsawat, Napassorn; Srionrod, Nitipon; Khumpim, Pacharaporn; Chawengkirttikul, Runglawan; Anuracpreeda, Panat

doi:10.1016/j.actatropica.2021.105933

Cited by 12 publications

(8 citation statements)

References 38 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Polyclonal anti-rCap protein was produced with the method described by Watthanadirek et al ( 18 , 19 ) and Junsiri et al ( 20 ). In brief, a female New Zealand White rabbit was injected with 500 μg of purified rCap protein mixed with 500 μL of complete Freund’s adjuvant to a final volume of 1,000 μL via a subcutaneous route.…”

Section: Methodsmentioning

confidence: 99%

Recombinant expression and characterization of Canine circovirus capsid protein for diagnosis

Dankaona,

Nooroong,

Poolsawat

et al. 2024

Front. Vet. Sci.

Self Cite

View full text Add to dashboard Cite

Canine circovirus (CanineCV) is a contagious virus that causes severe gastroenteritis, diarrhea, respiratory disease, and vasculitis, often resulting in fatality among infected dogs. In this study, a recombinant Capsid protein (rCap) of CanineCV was expressed in the Escherichia coli (E. coli) Rosetta (DE3) pLysS host cell, followed by affinity purification, and then analyzed by SDS-PAGE, revealing a molecular weight of approximately 31 kDa. The antigenicity of the CanineCV rCap protein was confirmed through recognition by a rabbit anti-CanineCV rCap protein polyclonal antibody (PoAb). Additionally, the reactivity and specificity of this PoAb were assessed using indirect enzyme-linked immunosorbent assay (ELISA) and Western blot analysis before applying in an immunohistochemistry (IHC), namely, immunoperoxidase detection. The immunoperoxidase assay using rabbit anti-CanineCV rCap protein PoAb demonstrated that the CanineCV Cap protein was predominantly located in immune cells, especially lymphocytes and macrophages, within the spleen, lung, tracheobronchial lymph nodes, small intestine, and kidney. Similarly, the Cap protein was also found in pneumocytes in the lung and renal tubular epithelial cells in the kidney. These findings reflected the biological activity and cell tropism of the virus. Therefore, the recombinant Cap protein and its PoAb could be used for the development of a valuable diagnostic tool for CanineCV detection.

show abstract

Section: Methodsmentioning

confidence: 99%

Recombinant expression and characterization of Canine circovirus capsid protein for diagnosis

Dankaona,

Nooroong,

Poolsawat

et al. 2024

Front. Vet. Sci.

Self Cite

View full text Add to dashboard Cite

show abstract

“…The genomic DNA of L. sabrazesi in blood samples was extracted by using an E.Z.N.A. ® Tissue DNA Kit (OMEGA Bio-Tek, Norcross, GA, USA) following the protocol of Watthanadirek et al [ 42 , 43 ] and Junsiri [ 22 ] with some modifications. Briefly, 250 μL of blood samples were mixed thoroughly with 25 μL of proteinase K solution, and incubated at 70 °C for 10 min.…”

Section: Methodsmentioning

confidence: 99%

Molecular genetic diversity and bioinformatic analysis of Leucocytozoon sabrazesi based on the mitochondrial genes cytb, coxI and coxIII and co-infection of Plasmodium spp.

et al. 2022

Self Cite

View full text Add to dashboard Cite

Leucocytozoon sabrazesi is an intracellular haemoprotozoan parasite responsible for leucocytozoonosis, which is transmitted by insect vectors and affects chickens in tropical and subtropical areas in many countries. It causes huge economic losses due to decreased meat and egg production. In the present study, we used nested PCR to determine the genetic diversity of L. sabrazesi based on the cytb, coxI, coxIII and concatenated genes in chickens in Thailand. In addition, we found co-infections between L. sabrazesi and Plasmodium spp. (P. gallinaceum or P. juxtanucleare) in chickens that were not identified by microscopic examination of blood smears. The phylogenetic analysis indicated that L. sabrazesi cytb and coxIII genes were conserved with similarity ranging from 99.9 to 100% and 98 to 100%, respectively whereas the coxI gene was diverse, with similarities ranging from 97 to 100%. These findings ascertained the nucleotide analysis of the cytb, coxI, coxIII and concatenated sequences in which 4, 8, 10 and 9 haplotypes were found, respectively. In addition, it was found that the large number of synonymous substitutions and conservative amino acid replacements in these mitochondrial genes occurred by non-synonymous substitution. The evolutionary analysis of the Ka/Ks ratio supported purifying selection and the negative values of both Fu’s Fs and Tajima’s D indicate selective sweep especially for the coxI gene. The entropy and Simplot analysis showed that the genetic variation in populations of Plasmodium spp. was higher than in Leucocytozoon. Hence, the nucleotide sequences of three mitochondrial genes could reflect the evolutionary analysis and geographic distribution of this protozoan population that switches hosts during its life cycle.

show abstract

Section: Methodsmentioning

confidence: 99%

“…Genomic DNA (gDNA) of B. bovis and B. bigemina was extracted from cattle blood samples using a Tissue DNA Extraction Kit (OMEGA, bio-tex, USA) following the methods described by Junsiri et al (2020) and Watthanadirek et al (2021) with some modifications. Briefly, 250 µl of blood sample was transferred into a 1.5 ml microcentrifuge tube, and 25 µl of Omega Bio-tek (OB) protease solution and 250 µl of BL buffer (lysis buffer) were added.…”

Section: Dna Extractionmentioning

confidence: 99%

Molecular characterization and genetic diversity of Babesia bovis and Babesia bigemina of cattle in Thailand

Srionrod

Nooroong

Poolsawat

et al. 2022

Front. Cell. Infect. Microbiol.

Self Cite

View full text Add to dashboard Cite

Babesia bovis and B. bigemina are the most common tick-borne parasites that cause bovine babesiosis which effects livestock production, leading to economic losses in tropical and subtropical areas of the world. The aims of this study were to determine the molecular detection, genetic diversity and antigenicity prediction of B. bovis based on spherical body protein 2 (sbp-2) gene and B. bigemina based on rhoptry-associated protein 1a (rap-1a) gene in cattle in Thailand. By PCR assay, the molecular detection of B. bovis and B. bigemina infection revealed levels of 2.58% (4/155) and 5.80% (9/155), respectively. The phylograms showed that B. bovis sbp-2 and B. bigemina rap-1a sequences displayed 5 and 3 clades with similarity ranging between 85.53 to 100% and 98.28 to 100%, respectively, when compared within Thailand strain. Diversity analysis of sbp-2 and rap-1a sequences showed 18 and 4 haplotypes, respectively. The entropy analysis illustrated 104 and 7 polymorphic sites of sbp-2 and rap-1a nucleic acid sequences, respectively, while those of sbp-2 and rap-1a amino acid sequences showed 46 and 4 high entropy peaks, respectively. Motifs analysis exhibited the distribution and conservation among sbp-2 and rap-1a sequences. The continuous and discontinuous B-cell epitopes have also been evaluated in this work. Therefore, our findings may be used to ameliorate the understanding inputs of molecular phylogeny, genetic diversity and antigenicity of B. bovis and B. bigemina Thailand stains.

show abstract

Molecular and recombinant characterization of major surface protein 5 from Anaplasma marginale

Cited by 12 publications

References 38 publications

Recombinant expression and characterization of Canine circovirus capsid protein for diagnosis

Recombinant expression and characterization of Canine circovirus capsid protein for diagnosis

Molecular genetic diversity and bioinformatic analysis of Leucocytozoon sabrazesi based on the mitochondrial genes cytb, coxI and coxIII and co-infection of Plasmodium spp.

Molecular characterization and genetic diversity of Babesia bovis and Babesia bigemina of cattle in Thailand

Contact Info

Product

Resources

About