Cloning, expression and partial characterization of a Haemaphysalis longicornis and a Rhipicephalus appendiculatus glutathione S‐transferase

Vaz, Itabajara da Silva; Imamura, Saiki; Ohashi, Kazuhiko; Onuma, Msayuki

doi:10.1111/j.0962-1075.2004.00493.x

Cited by 47 publications

(27 citation statements)

References 36 publications

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“…In this study, rHlYkt6/TrX rather than rHlYkt6/GST was employed to produce antibodies for the detection of native HlYkt6 protein in different tissues from ticks. This decision was based on the consideration that anti-rHlYkt6 sera developed from the rHlYkt6/GST protein can recognize the GST protein in tick tissues, since GST is ubiquitously expressed in ticks (da Silva Vaz Jnr et al, 2004).…”

Section: Discussionmentioning

confidence: 99%

Blocking the secretion of saliva by silencing the HlYkt6 gene in the tick Haemaphysalis longicornis

Gong¹,

Umemiya²,

Zhou³

et al. 2009

Insect Biochemistry and Molecular Biology

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Section: Discussionmentioning

confidence: 99%

Blocking the secretion of saliva by silencing the HlYkt6 gene in the tick Haemaphysalis longicornis

Gong¹,

Umemiya²,

Zhou³

et al. 2009

Insect Biochemistry and Molecular Biology

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“…The MSASignal fragment cloned in pCR TOPO 2.1 was digested with Bam HI (Invitrogen), purified and then ligated in plasmid pBlueScript ( pBS ) vector previously digested with same enzymes, and named pBlue-MSA plasmid. The HlGST sequence was amplified using HlGST-BamHI-SacII F and HlGST-PstI R primers (Table 2) using the plasmid pET43a-HlGST [30] as template. This PCR product was cloned in pCR TOPO 2.1, and termed p2 .…”

Section: Methodsmentioning

confidence: 99%

“…The anti-MSA1 monoclonal antibody BABB35 [39] was used as a positive control for the immunoblots at a concentration of 2μg/ml. Purified recombinant protein produced from pET43a-HlGST [30] was used for anti HlGST antibody production. One rabbit was inoculated four times at 15 days intervals by subcutaneous route with 100 μg of recombinant protein.…”

Section: Methodsmentioning

confidence: 99%

“…The use of GST in experimental vaccines resulted in variable degrees of protection against the targeted parasites, demonstrating their potential for generating protective immunity [29]. Furthermore, an experimental tick vaccine based on recombinant Haemaphysalis longicornis glutathione-S-transferase (HlGST) [30] elicited partially protective responses in bovines against R . microplus [21,22].…”

Section: Introductionmentioning

confidence: 99%

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Transfected Babesia bovis Expressing a Tick GST as a Live Vector Vaccine

et al. 2016

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The Rhipicephalus microplus tick is a notorious blood-feeding ectoparasite of livestock, especially cattle, responsible for massive losses in animal production. It is the main vector for transmission of pathogenic bacteria and parasites, including Babesia bovis, an intraerythrocytic apicomplexan protozoan parasite responsible for bovine Babesiosis. This study describes the development and testing of a live B. bovis vaccine expressing the protective tick antigen glutathione-S-transferase from Haemaphysalis longicornis (HlGST). The B. bovis S74-T3B parasites were electroporated with a plasmid containing the bidirectional Ef-1α (elongation factor 1 alpha) promoter of B. bovis controlling expression of two independent genes, the selectable marker GFP-BSD (green fluorescent protein–blasticidin deaminase), and HlGST fused to the MSA-1 (merozoite surface antigen 1) signal peptide from B. bovis. Electroporation followed by blasticidin selection resulted in the emergence of a mixed B. bovis transfected line (termed HlGST) in in vitro cultures, containing parasites with distinct patterns of insertion of both exogenous genes, either in or outside the Ef-1α locus. A B. bovis clonal line termed HlGST-Cln expressing intracellular GFP and HlGST in the surface of merozoites was then derived from the mixed parasite line HlGST using a fluorescent activated cell sorter. Two independent calf immunization trials were performed via intravenous inoculation of the HlGST-Cln and a previously described control consisting of an irrelevant transfected clonal line of B. bovis designated GFP-Cln. The control GFP-Cln line contains a copy of the GFP-BSD gene inserted into the Ef-1α locus of B. bovis in an identical fashion as the HIGST-Cln parasites. All animals inoculated with the HlGST-Cln and GFP-Cln transfected parasites developed mild babesiosis. Tick egg fertility and fully engorged female tick weight was reduced significantly in R. microplus feeding on HlGST-Cln-immunized calves. Collectively, these data show the efficacy of a transfected HlGST-Cln B. bovis parasite to induce detectable anti-glutathione-S-transferase antibodies and a reduction in tick size and fecundity of R. microplus feeding in experimentally inoculated animals.

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“…In ticks, however, the role of GSTs remains elusive. To date, four tick GSTs, two Boophilus microplus (Rosa de Lima et al ., 2002; He et al ., 1999), one Haemaphysalis longicornis and a Rhipicephalus appendiculatus (da Silva Vaz et al ., 2004) have been characterized.…”

Section: Introductionmentioning

confidence: 99%

Differential expression of two glutathione S‐transferases identified from the American dog tick,Dermacentor variabilis

Dreher-Lesnick

Mulenga

Simser

et al. 2006

Insect Molecular Biology

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Reciprocal signalling and gene expression play a cardinal role during pathogen-host molecular interactions and are prerequisite to the maintenance of balanced homeostasis. Gene expression repertoire changes during rickettsial infection and glutathione-S-transferases (GSTs) were among the genes found up-regulated in Rickettsia-infected Dermacentor variabilis. GSTs are well known to play an important part in cellular stress responses in the host. We have cloned two full-length GSTs from D. variabilis (DvGST1 and DvGST2). Comparison of these two DvGST molecules with those of other species indicate that DvGST1 is related to the mammalian class theta and insect class delta GSTs, while DvGST2 does not seem to fall in the same family. Northern blotting analyses revealed differential expression patterns, where DvGST1 and DvGST2 transcripts are found in the tick gut, with DvGST2 transcripts also present in the ovaries. Both DvGST transcripts are up-regulated upon tick feeding. Challenge of fed adult ticks with Escherichia coli injection showed decreased transcript amounts compared with ticks injected with phosphate-buffered saline (sham) and naïve ticks.

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Cloning, expression and partial characterization of a Haemaphysalis longicornis and a Rhipicephalus appendiculatus glutathione S‐transferase

Cited by 47 publications

References 36 publications

Blocking the secretion of saliva by silencing the HlYkt6 gene in the tick Haemaphysalis longicornis

Blocking the secretion of saliva by silencing the HlYkt6 gene in the tick Haemaphysalis longicornis

Transfected Babesia bovis Expressing a Tick GST as a Live Vector Vaccine

Differential expression of two glutathione S‐transferases identified from the American dog tick,Dermacentor variabilis

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