Full Text Available
Note: Clicking the button above will open the full text document at the original institutional repository in a new window.
A reverse genetics approach to evaluate Metzincins as anti-Rhipicephalus microplus tick vaccine candidates
Thesis (PhD)--University of Pretoria, 2013.
Saved in:
| Other Authors: | |
|---|---|
| Format: | Thesis |
| Language: | Eng |
| Published: |
University of Pretoria
2014
|
| Subjects: | |
| Tags: |
No Tags, Be the first to tag this record!
|
| _version_ | 1867613515625267200 |
|---|---|
| access_status_str | Open Access |
| author2 | Maritz-Olivier, Christine |
| author_browse | Maritz-Olivier, Christine |
| author_facet | Maritz-Olivier, Christine |
| collection | Thesis |
| dc_rights_str_mv | © 2013 University of Pretoria. All rights reserved. The copyright in this work vests in the University of Pretoria. No part of this work may be reproduced or transmitted in any form or by any means, without the prior written permission of the University of Pretoria |
| description | Thesis (PhD)--University of Pretoria, 2013. |
| format | Thesis |
| id | oai:repository.up.ac.za:2263/32964 |
| institution | University of Pretoria (South Africa) |
| language | Eng |
| last_indexed | 2026-06-10T12:37:22.799Z |
| license_str | Other — see source repository |
| provenance_str_mv | Harvested via OAI-PMH from UPSpace — University of Pretoria Institutional Repository |
| publishDate | 2014 |
| publishDateRange | 2014 |
| publishDateSort | 2014 |
| publisher | University of Pretoria |
| publisherStr | University of Pretoria |
| record_format | dspace |
| source_str | UPSpace — University of Pretoria Institutional Repository |
| spelling | oai:repository.up.ac.za:2263/32964 A reverse genetics approach to evaluate Metzincins as anti-Rhipicephalus microplus tick vaccine candidates Maritz-Olivier, Christine Neitz, A.W.H. (Albert Walter Herman) Gaspar, A.R.M. (Anabella Regina Marques) Barnard, Annette-Christi Vital physiological processes Anti-tick vaccines R. microplus Affect tick survival UCTD Thesis (PhD)--University of Pretoria, 2013. Tick proteins functioning in vital physiological processes such as blood meal uptake, digestion and reproduction are potential targets for anti-tick vaccines, since vaccination could disrupt these essential functions and ultimately affect tick survival. In this study we identified metzincin metalloproteases from R. microplus, the world’s most economically important external ectoparasite of cattle, as potential vaccine candidates since they are implicated to be essential to blood-cavity formation, bloodmeal digestion and reproduction in ixodid ticks. A vaccine derived from a single member of such a large family pose the obstacle of redundancy within the family, that may allow the function of the targeted family member to be taken up by other family members. Therefore the aim of this study was not only to focus on the physiological importance of each metzincin transcript, but also to investigate the differential gene expression network between the different metzincin family members. Eight transcripts encoding proteins containing the characteristic metzincin zinc-binding motif HEXXHXXG/NXXH/D and a unique methionine-turn were identified from native and in-house assembled R. microplus Expressed Sequence Tag databases. These were representative of five reprolysin-like and three astacin-like metzincin metalloproteases. Reverse transcription-PCR indicated that the reprolysins were most abundantly expressed in the salivary glands, whereas the astacins were most abundant in the midgut and ovaries. In vivo gene silencing utilizing RNA interference, was performed to assess a possible phenotype in silenced adult female R. microplus ticks during blood feeding and reproduction. RNAi against two reprolysins and one astacin significantly affected average egg weight as well as the oviposition rate. Moreover, integrated real time-PCR studies revealed an extensive cross organ network between the R. microplus metzincin transcripts, supporting the use of a combinatorial metzincinbased anti- R. microplus vaccine targeting multiple members of the large metzincin clan simultaneous. To conclusively evaluate the vaccination potential of the three identified metzincin candidates, the immunogenicity and protective properties of the recombinant proteins needs to be determined. Due to metalloproteases destructive activity and characteristics such as cysteine rich domains, only selected domains of the three candidates were expressed, using a cost effective Eshericia coli based expression system. Finally, the ability of each successfully expressed domain to elicit an immune response and serve as a protective antigen against R. microplus will be screened during vaccination trials in cattle. gm2013 Biochemistry unrestricted 2014-01-09T11:15:59Z 2014-01-09T11:15:59Z 2013-09-06 2013 Thesis Barnard, A. 2013, The role of behavioural aspects in investment decision-making by listed property fund managers in South Africa, PhD thesis, University of Pretoria, Pretoria, viewed yymmdd <http://hdl.handle.net/2263/32964> D13/9/883/gm http://hdl.handle.net/2263/32964 Eng © 2013 University of Pretoria. All rights reserved. The copyright in this work vests in the University of Pretoria. No part of this work may be reproduced or transmitted in any form or by any means, without the prior written permission of the University of Pretoria application/pdf University of Pretoria |
| spellingShingle | Vital physiological processes Anti-tick vaccines R. microplus Affect tick survival UCTD A reverse genetics approach to evaluate Metzincins as anti-Rhipicephalus microplus tick vaccine candidates |
| title | A reverse genetics approach to evaluate Metzincins as anti-Rhipicephalus microplus tick vaccine candidates |
| title_full | A reverse genetics approach to evaluate Metzincins as anti-Rhipicephalus microplus tick vaccine candidates |
| title_fullStr | A reverse genetics approach to evaluate Metzincins as anti-Rhipicephalus microplus tick vaccine candidates |
| title_full_unstemmed | A reverse genetics approach to evaluate Metzincins as anti-Rhipicephalus microplus tick vaccine candidates |
| title_short | A reverse genetics approach to evaluate Metzincins as anti-Rhipicephalus microplus tick vaccine candidates |
| title_sort | reverse genetics approach to evaluate metzincins as anti rhipicephalus microplus tick vaccine candidates |
| topic | Vital physiological processes Anti-tick vaccines R. microplus Affect tick survival UCTD |
| url | http://hdl.handle.net/2263/32964 |