A method to discriminate between closely related bovine major histocompatibility complex class I alleles by combining established PCR-SSP assays with RFLPs
We have developed a polymerase chain reaction-sequence-specific primers-restriction fragment length polymorphism (PCR-SSP-RFLP) method to rapidly differentiate between the A18 and A18 variant (v) BoLA haplotypes and between A14 and A15/A15v BoLA haplotypes in Holstein/Friesian cattle. We used publis...
| Autores principales: | , , , |
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| Formato: | Journal Article |
| Lenguaje: | Inglés |
| Publicado: |
Wiley
2015
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| Materias: | |
| Acceso en línea: | https://hdl.handle.net/10568/66014 |
| _version_ | 1855530678093873152 |
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| author | Svitek, Nicholas Nzau, Benjamin M. Steinaa, Lucilla Nene, Vishvanath M. |
| author_browse | Nene, Vishvanath M. Nzau, Benjamin M. Steinaa, Lucilla Svitek, Nicholas |
| author_facet | Svitek, Nicholas Nzau, Benjamin M. Steinaa, Lucilla Nene, Vishvanath M. |
| author_sort | Svitek, Nicholas |
| collection | Repository of Agricultural Research Outputs (CGSpace) |
| description | We have developed a polymerase chain reaction-sequence-specific primers-restriction fragment length polymorphism (PCR-SSP-RFLP) method to rapidly differentiate between the A18 and A18 variant (v) BoLA haplotypes and between A14 and A15/A15v BoLA haplotypes in Holstein/Friesian cattle. We used published SSP to PCR amplify BoLA alleles expressed in animals of known haplotype and exposed the amplicons to the restriction enzyme PvuII that was predicted to cut at a unique site in the middle of BoLA-6*01302 (A18v) and BoLA-1*00901(A15) but not in BoLA-6*01301 (A18) or BoLA-1*02301 (A14) alleles. Whereas the method does not discriminate between the A15 and A15v haplotypes, as the BoLA-1*00902 allele associated with A15v also contains a PvuII site, we are interested in cattle of A18 and A14 haplotype for vaccine related studies. Our results also indicated that the BoLA-6*01302 (A18v) allele is much more abundant than BoLA-6*01301 (A18) in the cattle that we sampled. |
| format | Journal Article |
| id | CGSpace66014 |
| institution | CGIAR Consortium |
| language | Inglés |
| publishDate | 2015 |
| publishDateRange | 2015 |
| publishDateSort | 2015 |
| publisher | Wiley |
| publisherStr | Wiley |
| record_format | dspace |
| spelling | CGSpace660142024-01-17T12:58:34Z A method to discriminate between closely related bovine major histocompatibility complex class I alleles by combining established PCR-SSP assays with RFLPs Svitek, Nicholas Nzau, Benjamin M. Steinaa, Lucilla Nene, Vishvanath M. theileria cattle animal diseases vaccines disease control east coast fever We have developed a polymerase chain reaction-sequence-specific primers-restriction fragment length polymorphism (PCR-SSP-RFLP) method to rapidly differentiate between the A18 and A18 variant (v) BoLA haplotypes and between A14 and A15/A15v BoLA haplotypes in Holstein/Friesian cattle. We used published SSP to PCR amplify BoLA alleles expressed in animals of known haplotype and exposed the amplicons to the restriction enzyme PvuII that was predicted to cut at a unique site in the middle of BoLA-6*01302 (A18v) and BoLA-1*00901(A15) but not in BoLA-6*01301 (A18) or BoLA-1*02301 (A14) alleles. Whereas the method does not discriminate between the A15 and A15v haplotypes, as the BoLA-1*00902 allele associated with A15v also contains a PvuII site, we are interested in cattle of A18 and A14 haplotype for vaccine related studies. Our results also indicated that the BoLA-6*01302 (A18v) allele is much more abundant than BoLA-6*01301 (A18) in the cattle that we sampled. 2015-04 2015-05-12T09:26:40Z 2015-05-12T09:26:40Z Journal Article https://hdl.handle.net/10568/66014 en Limited Access Wiley Svitek, N., Nzau, B., Steinaa, L. and Nene, V. 2015. A method to discriminate between closely related bovine major histocompatibility complex class I alleles by combining established PCR-SSP assays with RFLPs. Tissue Antigens 85(4):278-282. |
| spellingShingle | theileria cattle animal diseases vaccines disease control east coast fever Svitek, Nicholas Nzau, Benjamin M. Steinaa, Lucilla Nene, Vishvanath M. A method to discriminate between closely related bovine major histocompatibility complex class I alleles by combining established PCR-SSP assays with RFLPs |
| title | A method to discriminate between closely related bovine major histocompatibility complex class I alleles by combining established PCR-SSP assays with RFLPs |
| title_full | A method to discriminate between closely related bovine major histocompatibility complex class I alleles by combining established PCR-SSP assays with RFLPs |
| title_fullStr | A method to discriminate between closely related bovine major histocompatibility complex class I alleles by combining established PCR-SSP assays with RFLPs |
| title_full_unstemmed | A method to discriminate between closely related bovine major histocompatibility complex class I alleles by combining established PCR-SSP assays with RFLPs |
| title_short | A method to discriminate between closely related bovine major histocompatibility complex class I alleles by combining established PCR-SSP assays with RFLPs |
| title_sort | method to discriminate between closely related bovine major histocompatibility complex class i alleles by combining established pcr ssp assays with rflps |
| topic | theileria cattle animal diseases vaccines disease control east coast fever |
| url | https://hdl.handle.net/10568/66014 |
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