Thesis (MSc (Biochemistry))--University of Stellenbosch, 2010. / ENGLISH ABSTRACT:
The ostrich industry in South Africa is currently threatened by respiratory disease in feedlot ostriches
which causes a dramatic loss in production. Ms01, Ms02 and Ms03 were identified as the three ostrichspecific
mycoplasmas to be associated with this respiratory disease in ostriches of South Africa. The
ostrich-specific mycoplasmas have a major impact on ostrich production and for this reason there is a
serious need for treatment for these infections.
For this reason, the ostrich industry has undertaken an investigation into the development of vaccines
against mycoplasma infections. In this study, an approach to DNA vaccine development will be
investigated and applied, specifically for the ostrich mycoplasma Ms02. Firstly, the whole genome of
Ms02 was sequenced using GS FLX sequencing technology. The contiguous sequences obtained from the
whole-genome sequencing were analysed bioinformatically which included the annotation of the
contiguous sequences and the subsequent search for a vaccine candidate gene for the development of a
DNA vaccine. The P100 gene of Ms02, which showed a high degree of homology with the P100 gene of
the human pathogen M. hominis, was chosen as a vaccine candidate gene for the development of a DNA
vaccine. The P100 gene was successfully cloned and subsequently modified by means of site-directed
mutagenesis to correct for alternative codon usage, where after the modified P100 gene was subcloned
into the mammalian expression vector, pCI-neo for vaccination trials in the near future. / AFRIKAANSE OPSOMMING:
Die volstruisbedryf van Suid-Afrika is tans bedreig deur 'n respiratoriese siekte in voerkraal volstruise
wat lei tot aansienlike verliese in volstruisproduksie. Ms01, Ms02 en Ms03 is geïdentifiseer as die drie
volstruis-spesifieke mikoplasmas wat 'n rol speel in hierdie respiratoriese siektes van volstruise in Suid-
Afrika. Die drie volstruis-spesifieke mikoplasmas het 'n groot impak op die produksie van volstruise en
om hierdie rede is daar 'n ernstige behoefte aan 'n behandeling van hierdie infeksies.
Ten einde mikoplasma infeksies in volstruise te voorkom, het die Suid-Afrikaanse volstruisbedryf 'n
ondersoek geloods na moontlike strategieë vir entstof ontwikkeling. In hierdie studie, is 'n benadering van
DNA entstof ontwikkeling ondersoek en toegepas, spesifiek teen die volstruis mikoplasma Ms02.
Eerstens, is die volledige Ms02 genoomvolgorde bepaal deur gebruik te maak van GS FLX
volgordebepalingstegnologie. Die gedeeltelike volgordes verkry vanaf die heelgenoom volgordebepaling
is bioinformaties geanaliseer wat die annotering van die gedeeltelike volgordes asook die soektog vir 'n
kandidaat entstof geen vir die ontwikkeling van 'n DNA entstof ingesluit het. Die P100 geen van Ms02,
wat hoë homologie met die P100 geen van die menslike patogeen M. hominis getoon het, is gekies as die
kandidaat entstof geen. Die P100 geen is suksesvol gekloneer en gemodifiseer deur middel van setelgerigte
mutagenese om die P100 geen geskik te maak vir die invoeging in die soogdier ekspressie vektor,
pCI-neo vir toekomstige entstofproewe.
| Identifer | oai:union.ndltd.org:netd.ac.za/oai:union.ndltd.org:sun/oai:scholar.sun.ac.za:10019.1/5225 |
| Date | 12 1900 |
| Creators | Steenmans, Shandre |
| Contributors | Bellstedt, D. U., Botes, A., University of Stellenbosch. Faculty of Science. Dept. of Biochemistry. |
| Publisher | Stellenbosch : University of Stellenbosch |
| Source Sets | South African National ETD Portal |
| Language | en_ZA |
| Detected Language | English |
| Type | Thesis |
| Rights | University of Stellenbosch |
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