Verification of H5 and IL-15 gene expressions from H5-recombinant fowlpox viruses coexpressing host cytokine by using sds-page and western blotting
A development of effective vaccine as a treatment against avian influenza virus (AIV) has been introduced since the 1980s by using inactivated vaccines. A recombinant fowlpox virus expressing AIV haemagglutinin H5 gene, co-expressed with chicken interleukin (IL)-15 cytokine has been constructed prev...
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| Format: | Project Paper Report |
| Language: | English |
| Published: |
2015
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| Online Access: | http://psasir.upm.edu.my/id/eprint/90976/ http://psasir.upm.edu.my/id/eprint/90976/1/FBSB%202015%20164%20-%20IR.pdf |
| _version_ | 1848861217667416064 |
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| author | Zulkifli, Nurul Farhana |
| author_facet | Zulkifli, Nurul Farhana |
| author_sort | Zulkifli, Nurul Farhana |
| building | UPM Institutional Repository |
| collection | Online Access |
| description | A development of effective vaccine as a treatment against avian influenza virus (AIV) has been introduced since the 1980s by using inactivated vaccines. A recombinant fowlpox virus expressing AIV haemagglutinin H5 gene, co-expressed with chicken interleukin (IL)-15 cytokine has been constructed previously. In this study, the recombinant fowlpox virus (rFWPV) were analysed by using SDS-PAGE and Western blotting to verify the expression of H5 and IL-15 genes. The protein samples were separated by SDS-PAGE and transferred onto the surface of nitrocellulose membrane electrophoretically, by semi-dry Western blotting technique. Mouse β-actin, anti-IL-15, ab62587, and ab21292 primary antibodies were used to probe the membrane after blocking process. The antibody-antigen complexes were conjugated with alkaline phosphatase coupled to a secondary anti-IgG antibody which were either anti-mouse, anti-goat, or anti-rabbit, depending on the specific primary antibodies. The membranes were then developed by using commercially available WesternBreeze® Chromogenic Immunodetection System. Early findings revealed that false positive expressions were detected in all of the gel wells including negative controls, based on the presence of non-specific bands of 56 kD and 42 kD. Later results showed a 42 kD protein band size for β-actin expression, however the expressions of H5 and IL-15 proteins in the rFWPV were unsuccessful. |
| first_indexed | 2025-11-15T12:57:37Z |
| format | Project Paper Report |
| id | upm-90976 |
| institution | Universiti Putra Malaysia |
| institution_category | Local University |
| language | English |
| last_indexed | 2025-11-15T12:57:37Z |
| publishDate | 2015 |
| recordtype | eprints |
| repository_type | Digital Repository |
| spelling | upm-909762021-10-13T03:07:01Z http://psasir.upm.edu.my/id/eprint/90976/ Verification of H5 and IL-15 gene expressions from H5-recombinant fowlpox viruses coexpressing host cytokine by using sds-page and western blotting Zulkifli, Nurul Farhana A development of effective vaccine as a treatment against avian influenza virus (AIV) has been introduced since the 1980s by using inactivated vaccines. A recombinant fowlpox virus expressing AIV haemagglutinin H5 gene, co-expressed with chicken interleukin (IL)-15 cytokine has been constructed previously. In this study, the recombinant fowlpox virus (rFWPV) were analysed by using SDS-PAGE and Western blotting to verify the expression of H5 and IL-15 genes. The protein samples were separated by SDS-PAGE and transferred onto the surface of nitrocellulose membrane electrophoretically, by semi-dry Western blotting technique. Mouse β-actin, anti-IL-15, ab62587, and ab21292 primary antibodies were used to probe the membrane after blocking process. The antibody-antigen complexes were conjugated with alkaline phosphatase coupled to a secondary anti-IgG antibody which were either anti-mouse, anti-goat, or anti-rabbit, depending on the specific primary antibodies. The membranes were then developed by using commercially available WesternBreeze® Chromogenic Immunodetection System. Early findings revealed that false positive expressions were detected in all of the gel wells including negative controls, based on the presence of non-specific bands of 56 kD and 42 kD. Later results showed a 42 kD protein band size for β-actin expression, however the expressions of H5 and IL-15 proteins in the rFWPV were unsuccessful. 2015-06 Project Paper Report NonPeerReviewed text en http://psasir.upm.edu.my/id/eprint/90976/1/FBSB%202015%20164%20-%20IR.pdf Zulkifli, Nurul Farhana (2015) Verification of H5 and IL-15 gene expressions from H5-recombinant fowlpox viruses coexpressing host cytokine by using sds-page and western blotting. [Project Paper Report] |
| spellingShingle | Zulkifli, Nurul Farhana Verification of H5 and IL-15 gene expressions from H5-recombinant fowlpox viruses coexpressing host cytokine by using sds-page and western blotting |
| title | Verification of H5 and IL-15 gene expressions from H5-recombinant fowlpox viruses coexpressing host cytokine by using sds-page and western blotting |
| title_full | Verification of H5 and IL-15 gene expressions from H5-recombinant fowlpox viruses coexpressing host cytokine by using sds-page and western blotting |
| title_fullStr | Verification of H5 and IL-15 gene expressions from H5-recombinant fowlpox viruses coexpressing host cytokine by using sds-page and western blotting |
| title_full_unstemmed | Verification of H5 and IL-15 gene expressions from H5-recombinant fowlpox viruses coexpressing host cytokine by using sds-page and western blotting |
| title_short | Verification of H5 and IL-15 gene expressions from H5-recombinant fowlpox viruses coexpressing host cytokine by using sds-page and western blotting |
| title_sort | verification of h5 and il-15 gene expressions from h5-recombinant fowlpox viruses coexpressing host cytokine by using sds-page and western blotting |
| url | http://psasir.upm.edu.my/id/eprint/90976/ http://psasir.upm.edu.my/id/eprint/90976/1/FBSB%202015%20164%20-%20IR.pdf |