Overexpression and purification of YidRv gene from the hypervirulent Klebsiella pneumoniae, and the ability of the gene product in inducing a humoral response
The yidRv gene, isolated from the hypervirulent Klebsiella pneumoniae (hvKP), is a novel gene with an unknown function; however, it has exhibited high homology to the yidR, a gene recognized as potential vaccine candidate. The aim of this study was to clone the yidRv gene from the Indonesian hvKP an...
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Universitas Gadjah Mada, Yogyakarta
2024-12-01
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| Series: | Indonesian Journal of Biotechnology |
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| Online Access: | https://jurnal.ugm.ac.id/ijbiotech/article/view/95222 |
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| author | Tri Yudani Mardining Raras Mauludy Jutta Ajrullah Ellen Fenix Gunawan Nadya Shafa Pramesti Sumarno Reto Prawiro Agustin Krisna Wardani Is Helianti |
| author_facet | Tri Yudani Mardining Raras Mauludy Jutta Ajrullah Ellen Fenix Gunawan Nadya Shafa Pramesti Sumarno Reto Prawiro Agustin Krisna Wardani Is Helianti |
| author_sort | Tri Yudani Mardining Raras |
| collection | DOAJ |
| description | The yidRv gene, isolated from the hypervirulent Klebsiella pneumoniae (hvKP), is a novel gene with an unknown function; however, it has exhibited high homology to the yidR, a gene recognized as potential vaccine candidate. The aim of this study was to clone the yidRv gene from the Indonesian hvKP and to investigate its overexpression in Escherichia coli. In the experiment, yidRv was cloned into pET21 to construct pYik23. Recombinant protein YidRv was produced by growing E. coli BL21 (DE3)/pYik23 in LB medium with ampicillin at 29 °C, inducing protein synthesis with 0.5 mM IPTG for 20 hours. Purification was performed using Ni‐NTA resin, and the purified protein (50 µg) was administered to BALB/c mice to test for the production of IgG, IgM and IgA on 2 days before and day 19th and 37th after the first vaccination. The results show a significant induction of IgG and IgM, but not of IgA antibodies. In conclusion, the yidRv gene was overexpressed in E. coli BL21 (DE3) at high levels in soluble form, with the recombinant protein able to be purified to 90% homogeneity. The recombinant YidRv demonstrated the ability to stimulate the generation of both IgM and IgG antibodies. |
| format | Article |
| id | doaj-art-0117964563b544d0bbef8c6bbe84d422 |
| institution | DOAJ |
| issn | 0853-8654 2089-2241 |
| language | English |
| publishDate | 2024-12-01 |
| publisher | Universitas Gadjah Mada, Yogyakarta |
| record_format | Article |
| series | Indonesian Journal of Biotechnology |
| spelling | doaj-art-0117964563b544d0bbef8c6bbe84d4222025-08-20T03:08:18ZengUniversitas Gadjah Mada, YogyakartaIndonesian Journal of Biotechnology0853-86542089-22412024-12-0129419820410.22146/ijbiotech.9522237006Overexpression and purification of YidRv gene from the hypervirulent Klebsiella pneumoniae, and the ability of the gene product in inducing a humoral responseTri Yudani Mardining Raras0Mauludy Jutta Ajrullah1Ellen Fenix Gunawan2Nadya Shafa Pramesti3Sumarno Reto Prawiro4Agustin Krisna Wardani5Is Helianti6Department of Biochemistry‐Molecular Biology, Faculty of Medicine, Universitas Brawijaya, Jl. Veteran, Malang, IndonesiaMaster Program in Biomedical Science, Faculty of Medicine, Universitas Brawijaya, Jl. Veteran, Malang, IndonesiaBachelor Program in Biotechnology, Faculty of Agricultural Technology, Universitas Brawijaya, Jl. Veteran, Malang, IndonesiaBachelor Program in Biotechnology, Faculty of Agricultural Technology, Universitas Brawijaya, Jl. Veteran, Malang, IndonesiaDepartment of Clinical Microbiology, Faculty of Medicine, Universitas Brawijaya, Jl. Veteran, Malang, IndonesiaDepartment of Food Science and Biotechnology, Faculty of Agricultural Technology, Universitas Brawijaya, Jl. Veteran, Malang, IndonesiaResearch Center for Genetic Engineering, KST Soekarno, Jalan Raya Bogor Km 46, Cibinong, Jawa Barat, IndonesiaThe yidRv gene, isolated from the hypervirulent Klebsiella pneumoniae (hvKP), is a novel gene with an unknown function; however, it has exhibited high homology to the yidR, a gene recognized as potential vaccine candidate. The aim of this study was to clone the yidRv gene from the Indonesian hvKP and to investigate its overexpression in Escherichia coli. In the experiment, yidRv was cloned into pET21 to construct pYik23. Recombinant protein YidRv was produced by growing E. coli BL21 (DE3)/pYik23 in LB medium with ampicillin at 29 °C, inducing protein synthesis with 0.5 mM IPTG for 20 hours. Purification was performed using Ni‐NTA resin, and the purified protein (50 µg) was administered to BALB/c mice to test for the production of IgG, IgM and IgA on 2 days before and day 19th and 37th after the first vaccination. The results show a significant induction of IgG and IgM, but not of IgA antibodies. In conclusion, the yidRv gene was overexpressed in E. coli BL21 (DE3) at high levels in soluble form, with the recombinant protein able to be purified to 90% homogeneity. The recombinant YidRv demonstrated the ability to stimulate the generation of both IgM and IgG antibodies.https://jurnal.ugm.ac.id/ijbiotech/article/view/95222hypervirulentklebsiella pneumoniaeoverexpressionrecombinant antigen yidrv |
| spellingShingle | Tri Yudani Mardining Raras Mauludy Jutta Ajrullah Ellen Fenix Gunawan Nadya Shafa Pramesti Sumarno Reto Prawiro Agustin Krisna Wardani Is Helianti Overexpression and purification of YidRv gene from the hypervirulent Klebsiella pneumoniae, and the ability of the gene product in inducing a humoral response Indonesian Journal of Biotechnology hypervirulent klebsiella pneumoniae overexpression recombinant antigen yidrv |
| title | Overexpression and purification of YidRv gene from the hypervirulent Klebsiella pneumoniae, and the ability of the gene product in inducing a humoral response |
| title_full | Overexpression and purification of YidRv gene from the hypervirulent Klebsiella pneumoniae, and the ability of the gene product in inducing a humoral response |
| title_fullStr | Overexpression and purification of YidRv gene from the hypervirulent Klebsiella pneumoniae, and the ability of the gene product in inducing a humoral response |
| title_full_unstemmed | Overexpression and purification of YidRv gene from the hypervirulent Klebsiella pneumoniae, and the ability of the gene product in inducing a humoral response |
| title_short | Overexpression and purification of YidRv gene from the hypervirulent Klebsiella pneumoniae, and the ability of the gene product in inducing a humoral response |
| title_sort | overexpression and purification of yidrv gene from the hypervirulent klebsiella pneumoniae and the ability of the gene product in inducing a humoral response |
| topic | hypervirulent klebsiella pneumoniae overexpression recombinant antigen yidrv |
| url | https://jurnal.ugm.ac.id/ijbiotech/article/view/95222 |
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