Valosin-containing protein from the hard tick Haemaphysalis longicornis: effects of dsRNA-mediated HlVCP gene silencing

2007 ◽  
Vol 85 (3) ◽  
pp. 384-394 ◽  
Author(s):  
Damdinsuren Boldbaatar ◽  
Badgar Battsetseg ◽  
Takeshi Hatta ◽  
Takeharu Miyoshi ◽  
Naotoshi Tsuji ◽  
...  
Keyword(s):  
Blood Feeding ◽  
Pcr Analysis ◽  
Hard Tick ◽  
Haemaphysalis Longicornis ◽  
Native Protein ◽  
Rt Pcr ◽  
Future Studies ◽  
Atpase Gene ◽  
A Domains

We report the cloning and characterization of a cDNA encoding the valosin-containing protein (VCP) from the Haemaphysalis longicornis tick (HlVCP). The full-length HlVCP is 2782 bp and codes for 808 amino acids of a deduced protein with a predicted molecular mass of 89.9 kDa. The domain structure analysis revealed that the deduced protein has 2 Walker A domains, 2 Walker B domains, a Cdc48 domain, and a polyQ-binding domain. The mouse anti-HlVCP serum recognized a 97 kDa native protein in the salivary glands, midgut, and synganglion. RT–PCR analysis revealed that the native VCP was expressed throughout the developing stages and in tick organs. HlVCP silencing resulted in a decrease in tick body mass after blood feeding. This study not only contributes to a growing understanding of the ATPase gene family but also lays the groundwork for future studies on protein secretion and host–tick interaction. This study is the first report of the VCP gene from Chelicerata, which include spiders, scorpions, and ticks.

Identification of a gene cluster for cell-surface genes of the SRS superfamily inNeospora caninumand characterization of the novelSRS9gene

Parasitology ◽  
2011 ◽  
Vol 138 (14) ◽  
pp. 1832-1842 ◽  
Author(s):  
V. RISCO-CASTILLO ◽  
V. MARUGÁN-HERNÁNDEZ ◽  
A. FERNÁNDEZ-GARCÍA ◽  
A. AGUADO-MARTÍNEZ ◽  
E. JIMÉNEZ-RUIZ ◽  
...  
Keyword(s):  
Western Blot ◽  
Gene Cluster ◽  
Neospora Caninum ◽  
Pcr Analysis ◽  
Rt Pcr ◽  
Specific Expression ◽  
Initial Characterization ◽  
Hydrophilic Region

SUMMARYHere we present the detection of a gene cluster forNeospora caninumsurface genes, similar to theToxoplasma gondiiSRS9 locus, and the cloning and characterization of the NcSRS9gene. PCR genome walking, using NcBSR4gene as a framework, allows the identification, upstream NcBSR4, of 2 sequences homologous to theSRS5and the Ubiquinol-cytochrome C reductase genes and, downstream NcBSR4, of an ORF of 1191 bp coding for a 396-amino acid polypeptide with 59% similarity to the TgSRS9 antigen. A putative 39-residue signal peptide was found at the NH2-terminus followed by a hydrophilic region, and a potential site for a glycosylphosphatidylinositol anchor at the COOH-terminus. A recombinant NcSRS9 protein was produced and was recognized on a Western blot by a low proportion of sera from a panel of naturally infected cows and calves. In addition, Western blot analysis using polyclonal anti-rNcSRS9 revealed stage-specific expression of NcSRS9 in bradyzoites but not in tachyzoites, and immunohistochemistry on brain from a congenitally infected calf showed NcSRS9 recognition in bradyzoites contained in tissue cysts. However, bradyzoite-specific expression of NcSRS9 could not be proven by immunofluorescence on bradyzoites obtainedin vitroand RT-PCR analysis showed no significant variations of NcSRS9transcripts duringin vitrotachyzoite-bradyzoite switch, probably due to incomplete maturity ofin vitrobradyzoites. Initial characterization of NcSRS9 in this study may lead to further studies for a better understanding ofN. caninumpersistence.

Molecular cloning and functional characterization of an aspartic protease from the hard tick Haemaphysalis longicornis

2006 ◽  
Vol 36 (1) ◽  
pp. 25-36 ◽  
Author(s):  
Damdinsuren Boldbaatar ◽  
Chummy Sikalizyo Sikasunge ◽  
Badgar Battsetseg ◽  
Xuenan Xuan ◽  
Kozo Fujisaki
Keyword(s):  
Molecular Cloning ◽  
Functional Characterization ◽  
Aspartic Protease ◽  
Hard Tick ◽  
Haemaphysalis Longicornis

Cloning and molecular characterization of a cubilin-related serine proteinase from the hard tick Haemaphysalis longicornis

2004 ◽  
Vol 34 (8) ◽  
pp. 799-808 ◽  
Author(s):  
Takeharu Miyoshi ◽  
Naotoshi Tsuji ◽  
M Khyrul Islam ◽  
Tsugihiko Kamio ◽  
Kozo Fujisaki
Keyword(s):  
Molecular Characterization ◽  
Serine Proteinase ◽  
Hard Tick ◽  
Haemaphysalis Longicornis

scholarly journals Enzymatic Characterization of a Cubilin-Related Serine Proteinase from the Hard Tick Haemaphysalis longicornis

2004 ◽  
Vol 66 (10) ◽  
pp. 1195-1198 ◽  
Author(s):  
Takeharu MIYOSHI ◽  
Naotoshi TSUJI ◽  
M. Khyrul ISLAM ◽  
Tsugihiko KAMIO ◽  
Kozo FUJISAKI
Keyword(s):  
Serine Proteinase ◽  
Hard Tick ◽  
Haemaphysalis Longicornis ◽  
Enzymatic Characterization

Naked Poly(amidoamine) Dendrimer Nanoparticles Exhibit Intrinsic Embryotoxicity During the Early Stages of Normal Development

2020 ◽  
Vol 16 (10) ◽  
pp. 1454-1462
Author(s):  
Hadeel Kheraldine ◽  
Ishita Gupta ◽  
Hashim Alhussain ◽  
Ayesha Jabeen ◽  
Saghir Akhtar ◽  
...  
Keyword(s):  
Normal Development ◽  
Chorioallantoic Membrane ◽  
Pamam Dendrimers ◽  
Pcr Analysis ◽  
Dependent Manner ◽  
Rt Pcr ◽  
Future Studies ◽  
Early Stages ◽  
Expression Of Genes ◽  
The Impact

To investigate the impact of poly(amidoamine) dendrimers (PAMAMs) in the embryo, we explored the outcome of different generations (G4 and G6) on the early stages of embryogenesis using the chicken embryo as a model. We also monitored their effect on angiogenesis in the chorioallantoic membrane (CAM). Our data revealed that cationic PAMAMs provoke substantial embryotoxicity, as they significantly induce death (up to 50%, p < 0 05) and inhibit angiogenesis of the CAM (up to 30%, p < 0 05) in a generation-dependent manner in comparison to controls and other types of PAMAMs (anionic and neutral). Moreover, cationic PAMAMs alter the expression of genes related to cell survival, cell cycle, proliferation, transcription factor, apoptosis, and angiogenesis, as shown by RT-PCR analysis. Our data suggest that PAMAM dendrimers exhibit intrinsic toxicity in embryos at the early stages and inhibits angiogenesis of the CAM. Thus, future studies are necessary to illustrate the exact mechanism of PAMAM dendrimers in embryotoxicity.

scholarly journals The identification and characterization of a testis-specific cDNA during spermatogenesis

2006 ◽  
Vol 11 (1) ◽  
Author(s):  
Ying Chen ◽  
Jiarui Hu ◽  
Ping Song ◽  
Wuming Gong
Keyword(s):  
Experimental Validation ◽  
Rna Binding ◽  
Pcr Analysis ◽  
Splicing Factors ◽  
Rt Pcr ◽  
Rich Domain ◽  
Pachytene Spermatocytes ◽  
Identification And Characterization

AbstractUsing bioinformatics and experimental validation, we obtained a cDNA (named srsf) which was exclusively expressed in the mouse testes. RT-PCR analysis showed that srsf mRNA was not expressed in the gonad during the sex determination period or during embryogenesis. In developing mouse tests, srsf expression was first detected on post-natal day 10, reached its highest level on day 23, and then reduced to and remained at a moderate level throughout adulthood. In situ hybridization analysis demonstrated that srsf mRNA was expressed in pachytene spermatocytes and round spermatids in the testes. The predicted protein contains one RNA-binding domain (RBD) and a serine-arginine rich domain (RS), which are characterized by some splicing factors of SR family members. These findings indicate that srsf may play a role during spermatogenesis.

Legumains from the hard tick Haemaphysalis longicornis play modulatory roles in blood feeding and gut cellular remodelling and impact on embryogenesis

2009 ◽  
Vol 39 (1) ◽  
pp. 97-107 ◽  
Author(s):  
M. Abdul Alim ◽  
Naotoshi Tsuji ◽  
Takeharu Miyoshi ◽  
M. Khyrul Islam ◽  
Takeshi Hatta ◽  
...  
Keyword(s):  
Blood Feeding ◽  
Hard Tick ◽  
Haemaphysalis Longicornis

scholarly journals EB8 Molecular characterization of a peroxiredoxin from the hard tick Haemaphysalis longicornis

2001 ◽  
Vol 52 (Supplement) ◽  
pp. 141
Author(s):  
Naotoshi TSUJI ◽  
Myung Jo YOO ◽  
Harue KASUGA ◽  
Takashi ISOBE ◽  
Tsugihiko KAMIO ◽  
...  
Keyword(s):  
Molecular Characterization ◽  
Hard Tick ◽  
Haemaphysalis Longicornis

Molecular cloning and characterization of gut-derived cysteine proteinases associated with a host protective extract from Haemonchus contortus

Parasitology ◽  
1999 ◽  
Vol 119 (4) ◽  
pp. 405-412 ◽  
Author(s):  
P. J. SKUCE ◽  
D. L. REDMOND ◽  
S. LIDDELL ◽  
E. M. STEWART ◽  
G. F. J. NEWLANDS ◽  
...  
Keyword(s):  
Haemonchus Contortus ◽  
Southern Blot Analysis ◽  
Single Copy ◽  
Blood Feeding ◽  
Cdna Expression Library ◽  
Cysteine Proteinases ◽  
Rt Pcr ◽  
Expression Library ◽  
Developmentally Regulated

Cysteine proteinases have been implicated in the protection conferred by vaccination with detergent-soluble extracts of Haemonchus contortus. In the present study, antisera from sheep refractory to Haemonchus challenge following vaccination with a ‘proteinase-enriched’ Haemonchus gut membrane extract, were employed to screen a cDNA expression library of the adult parasite. This resulted in the isolation of 3 cDNAs (designated hmcp1, 4 and 6) encoding cathepsin B-like cysteine proteinases. Immunocytochemical studies specifically localized the products of these genes to the microvillar surface of the parasite's gut and RT–PCR experiments revealed that these were developmentally regulated, being expressed exclusively during the blood-feeding parasitic stages. In addition, a generic PCR approach was adopted in order to identify the predominant cysteine proteinases in a UK strain of Haemonchus. A panel of 5 cDNAs, including hmcp1 and 4, was amplified in this way. Genomic Southern blot analysis indicated that some of these enzymes were encoded by single-copy genes, whereas others were encoded by multi-copy genes. Subsequent sequence analysis revealed that the proteases identified in this study were distinct from those previously reported in USA strains of the parasite.

scholarly journals Functional characterization of two defensins, HlDFS1 and HlDFS2, from the hard tick Haemaphysalis longicornis

2017 ◽  
Vol 10 (1) ◽  
Author(s):  
Ta Sun ◽  
Wen Pan ◽  
Yanhui Song ◽  
Jingpin Zhang ◽  
Jingwen Wang ◽  
...  
Keyword(s):  
Functional Characterization ◽  
Hard Tick ◽  
Haemaphysalis Longicornis
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