Molecular Cloning and Functional Characterization of the 5′ Regulatory Region of the <i>SLC11A1</i> Gene from Yaks
The solute transport protein family 11 A1 (<i>SLC11A1</i>), also recognized as natural resistance-associated macrophage protein 1 (<i>NRAMP1</i>), represents a transmembrane protein encoded by the <i>SLC11A1</i> gene. A variety of prior investigations have illumin...
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MDPI AG
2023-11-01
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| Series: | Animals |
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| Online Access: | https://www.mdpi.com/2076-2615/13/23/3615 |
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| author | Yuqing Chong Liping Wang Bo Wang Zhendong Gao Ying Lu Weidong Deng Dongmei Xi |
| author_facet | Yuqing Chong Liping Wang Bo Wang Zhendong Gao Ying Lu Weidong Deng Dongmei Xi |
| author_sort | Yuqing Chong |
| collection | DOAJ |
| description | The solute transport protein family 11 A1 (<i>SLC11A1</i>), also recognized as natural resistance-associated macrophage protein 1 (<i>NRAMP1</i>), represents a transmembrane protein encoded by the <i>SLC11A1</i> gene. A variety of prior investigations have illuminated its involvement in conferring resistance or susceptibility to bacterial agents, positioning it as a promising candidate gene for breeding disease-resistant animals. Yaks (<i>Bos grunniens</i>), renowned inhabitants of the Qinghai-Tibet Plateau in China, stand as robust ruminants distinguished by their adaptability and formidable disease resistance. Notwithstanding these unique traits, there is scant literature on the <i>SLC11A1</i> gene in the yak population. Our inquiry commences with the cloning of the 5′ regulatory region sequence of the Zhongdian yak <i>SLC11A1</i> gene. We employ bioinformatics tools to identify transcription factor binding sites, delineating pivotal elements like enhancers and cis-acting elements. To ascertain the promoter activity of this region, we amplify four distinct promoter fragments within the 5′ regulatory region of the yak <i>SLC11A1</i> gene. Subsequently, we design a luciferase reporter gene vector containing four site-specific deletion mutations and perform transient transfection experiments. Through these experiments, we measure and compare the activity of disparate gene fragments located within the 5′ regulatory region, revealing regions bearing promoter functionality and discerning key regulatory elements. Our findings validate the promoter functionality of the 5′ regulatory region, offering preliminary insights into the core and principal regulatory segments of this promoter. Notably, we identified single nucleotide polymorphisms (SNPs) that may be associated with important regulatory elements such as NF-1 and NF-1/L. This study provides a theoretical framework for in-depth research on the function and expression regulation mechanism of the yak <i>SLC11A1</i> gene. |
| first_indexed | 2024-03-09T01:56:27Z |
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| institution | Directory Open Access Journal |
| issn | 2076-2615 |
| language | English |
| last_indexed | 2024-03-09T01:56:27Z |
| publishDate | 2023-11-01 |
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| record_format | Article |
| series | Animals |
| spelling | doaj.art-4a852743c7a9498cab7da3336bb9b2fe2023-12-08T15:10:27ZengMDPI AGAnimals2076-26152023-11-011323361510.3390/ani13233615Molecular Cloning and Functional Characterization of the 5′ Regulatory Region of the <i>SLC11A1</i> Gene from YaksYuqing Chong0Liping Wang1Bo Wang2Zhendong Gao3Ying Lu4Weidong Deng5Dongmei Xi6Faculty of Animal Science and Technology, Yunnan Agricultural University, Kunming 650201, ChinaFaculty of Biological Engineering, Xinxiang University, Xinxiang 453003, ChinaFaculty of Animal Science and Technology, Yunnan Agricultural University, Kunming 650201, ChinaFaculty of Animal Science and Technology, Yunnan Agricultural University, Kunming 650201, ChinaFaculty of Animal Science and Technology, Yunnan Agricultural University, Kunming 650201, ChinaFaculty of Animal Science and Technology, Yunnan Agricultural University, Kunming 650201, ChinaFaculty of Animal Science and Technology, Yunnan Agricultural University, Kunming 650201, ChinaThe solute transport protein family 11 A1 (<i>SLC11A1</i>), also recognized as natural resistance-associated macrophage protein 1 (<i>NRAMP1</i>), represents a transmembrane protein encoded by the <i>SLC11A1</i> gene. A variety of prior investigations have illuminated its involvement in conferring resistance or susceptibility to bacterial agents, positioning it as a promising candidate gene for breeding disease-resistant animals. Yaks (<i>Bos grunniens</i>), renowned inhabitants of the Qinghai-Tibet Plateau in China, stand as robust ruminants distinguished by their adaptability and formidable disease resistance. Notwithstanding these unique traits, there is scant literature on the <i>SLC11A1</i> gene in the yak population. Our inquiry commences with the cloning of the 5′ regulatory region sequence of the Zhongdian yak <i>SLC11A1</i> gene. We employ bioinformatics tools to identify transcription factor binding sites, delineating pivotal elements like enhancers and cis-acting elements. To ascertain the promoter activity of this region, we amplify four distinct promoter fragments within the 5′ regulatory region of the yak <i>SLC11A1</i> gene. Subsequently, we design a luciferase reporter gene vector containing four site-specific deletion mutations and perform transient transfection experiments. Through these experiments, we measure and compare the activity of disparate gene fragments located within the 5′ regulatory region, revealing regions bearing promoter functionality and discerning key regulatory elements. Our findings validate the promoter functionality of the 5′ regulatory region, offering preliminary insights into the core and principal regulatory segments of this promoter. Notably, we identified single nucleotide polymorphisms (SNPs) that may be associated with important regulatory elements such as NF-1 and NF-1/L. This study provides a theoretical framework for in-depth research on the function and expression regulation mechanism of the yak <i>SLC11A1</i> gene.https://www.mdpi.com/2076-2615/13/23/3615Zhongdian yak<i>SLC11A1</i> gene5′ regulatory regionluciferase reporter genetransient transfection |
| spellingShingle | Yuqing Chong Liping Wang Bo Wang Zhendong Gao Ying Lu Weidong Deng Dongmei Xi Molecular Cloning and Functional Characterization of the 5′ Regulatory Region of the <i>SLC11A1</i> Gene from Yaks Animals Zhongdian yak <i>SLC11A1</i> gene 5′ regulatory region luciferase reporter gene transient transfection |
| title | Molecular Cloning and Functional Characterization of the 5′ Regulatory Region of the <i>SLC11A1</i> Gene from Yaks |
| title_full | Molecular Cloning and Functional Characterization of the 5′ Regulatory Region of the <i>SLC11A1</i> Gene from Yaks |
| title_fullStr | Molecular Cloning and Functional Characterization of the 5′ Regulatory Region of the <i>SLC11A1</i> Gene from Yaks |
| title_full_unstemmed | Molecular Cloning and Functional Characterization of the 5′ Regulatory Region of the <i>SLC11A1</i> Gene from Yaks |
| title_short | Molecular Cloning and Functional Characterization of the 5′ Regulatory Region of the <i>SLC11A1</i> Gene from Yaks |
| title_sort | molecular cloning and functional characterization of the 5 regulatory region of the i slc11a1 i gene from yaks |
| topic | Zhongdian yak <i>SLC11A1</i> gene 5′ regulatory region luciferase reporter gene transient transfection |
| url | https://www.mdpi.com/2076-2615/13/23/3615 |
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