Cloning, Expression, and Functional Characterization of FUT1, a Key Gene for Histo-Blood Group Antigens Synthesis in <i>Crassostrea gigas</i>
Histo-blood group antigens (HBGAs) comprise a family of cell-surface carbohydrates that are considered norovirus-specific binding receptors or ligands. HBGA-like molecules have also been detected in oysters as common norovirus carriers, although the pathway involved in the synthesis of these molecul...
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MDPI AG
2023-05-01
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| Series: | Current Issues in Molecular Biology |
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| author | Binbin Gui Lin Yao Meng Qu Weiran Zhang Mingyu Li Yanhua Jiang Lianzhu Wang |
| author_facet | Binbin Gui Lin Yao Meng Qu Weiran Zhang Mingyu Li Yanhua Jiang Lianzhu Wang |
| author_sort | Binbin Gui |
| collection | DOAJ |
| description | Histo-blood group antigens (HBGAs) comprise a family of cell-surface carbohydrates that are considered norovirus-specific binding receptors or ligands. HBGA-like molecules have also been detected in oysters as common norovirus carriers, although the pathway involved in the synthesis of these molecules in oysters has yet to be elucidated. We isolated and identified a key gene involved in the synthesis of HBGA-like molecules, <i>FUT1</i>, from <i>Crassostrea gigas</i>, named <i>CgFUT1</i>. Real-time quantitative polymerase chain reaction analysis showed that <i>CgFUT1</i> mRNA was expressed in the mantle, gill, muscle, labellum, and hepatopancreatic tissues of <i>C. gigas</i>, with the hepatopancreas exhibiting the highest expression level. A recombinant CgFUT1 protein with a molecular mass of 38.0 kDa was expressed in <i>Escherichia coli</i> using a prokaryotic expression vector. A eukaryotic expression plasmid was constructed and transfected into Chinese hamster ovary (CHO) cells. The expression of CgFUT1 and membrane localization of type H-2 HBGA-like molecules in CHO cells were detected using Western blotting and cellular immunofluorescence, respectively. This study indicated that <i>CgFUT1</i>, expressed in <i>C. gigas</i> tissues, can synthesize type H-2 HBGA-like molecules. This finding provides a new perspective for analyzing the source and synthetic pathway of HBGA-like molecules in oysters. |
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| issn | 1467-3037 1467-3045 |
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| spelling | doaj.art-df1c9e854b31431b8ecb92f4b31bce3a2023-11-18T00:57:08ZengMDPI AGCurrent Issues in Molecular Biology1467-30371467-30452023-05-014554200421310.3390/cimb45050267Cloning, Expression, and Functional Characterization of FUT1, a Key Gene for Histo-Blood Group Antigens Synthesis in <i>Crassostrea gigas</i>Binbin Gui0Lin Yao1Meng Qu2Weiran Zhang3Mingyu Li4Yanhua Jiang5Lianzhu Wang6School of Food Science and Technology, Dalian Polytechnic University, Dalian 116034, ChinaKey Laboratory of Testing and Evaluation for Aquatic Product Safety and Quality, Ministry of Agriculture and Rural Affairs, Qingdao 266071, ChinaKey Laboratory of Testing and Evaluation for Aquatic Product Safety and Quality, Ministry of Agriculture and Rural Affairs, Qingdao 266071, ChinaKey Laboratory of Testing and Evaluation for Aquatic Product Safety and Quality, Ministry of Agriculture and Rural Affairs, Qingdao 266071, ChinaKey Laboratory of Testing and Evaluation for Aquatic Product Safety and Quality, Ministry of Agriculture and Rural Affairs, Qingdao 266071, ChinaKey Laboratory of Testing and Evaluation for Aquatic Product Safety and Quality, Ministry of Agriculture and Rural Affairs, Qingdao 266071, ChinaSchool of Food Science and Technology, Dalian Polytechnic University, Dalian 116034, ChinaHisto-blood group antigens (HBGAs) comprise a family of cell-surface carbohydrates that are considered norovirus-specific binding receptors or ligands. HBGA-like molecules have also been detected in oysters as common norovirus carriers, although the pathway involved in the synthesis of these molecules in oysters has yet to be elucidated. We isolated and identified a key gene involved in the synthesis of HBGA-like molecules, <i>FUT1</i>, from <i>Crassostrea gigas</i>, named <i>CgFUT1</i>. Real-time quantitative polymerase chain reaction analysis showed that <i>CgFUT1</i> mRNA was expressed in the mantle, gill, muscle, labellum, and hepatopancreatic tissues of <i>C. gigas</i>, with the hepatopancreas exhibiting the highest expression level. A recombinant CgFUT1 protein with a molecular mass of 38.0 kDa was expressed in <i>Escherichia coli</i> using a prokaryotic expression vector. A eukaryotic expression plasmid was constructed and transfected into Chinese hamster ovary (CHO) cells. The expression of CgFUT1 and membrane localization of type H-2 HBGA-like molecules in CHO cells were detected using Western blotting and cellular immunofluorescence, respectively. This study indicated that <i>CgFUT1</i>, expressed in <i>C. gigas</i> tissues, can synthesize type H-2 HBGA-like molecules. This finding provides a new perspective for analyzing the source and synthetic pathway of HBGA-like molecules in oysters.https://www.mdpi.com/1467-3045/45/5/267<i>Crassostrea gigas</i>norovirusHBGAsgene expressionCgFUT1 |
| spellingShingle | Binbin Gui Lin Yao Meng Qu Weiran Zhang Mingyu Li Yanhua Jiang Lianzhu Wang Cloning, Expression, and Functional Characterization of FUT1, a Key Gene for Histo-Blood Group Antigens Synthesis in <i>Crassostrea gigas</i> Current Issues in Molecular Biology <i>Crassostrea gigas</i> norovirus HBGAs gene expression CgFUT1 |
| title | Cloning, Expression, and Functional Characterization of FUT1, a Key Gene for Histo-Blood Group Antigens Synthesis in <i>Crassostrea gigas</i> |
| title_full | Cloning, Expression, and Functional Characterization of FUT1, a Key Gene for Histo-Blood Group Antigens Synthesis in <i>Crassostrea gigas</i> |
| title_fullStr | Cloning, Expression, and Functional Characterization of FUT1, a Key Gene for Histo-Blood Group Antigens Synthesis in <i>Crassostrea gigas</i> |
| title_full_unstemmed | Cloning, Expression, and Functional Characterization of FUT1, a Key Gene for Histo-Blood Group Antigens Synthesis in <i>Crassostrea gigas</i> |
| title_short | Cloning, Expression, and Functional Characterization of FUT1, a Key Gene for Histo-Blood Group Antigens Synthesis in <i>Crassostrea gigas</i> |
| title_sort | cloning expression and functional characterization of fut1 a key gene for histo blood group antigens synthesis in i crassostrea gigas i |
| topic | <i>Crassostrea gigas</i> norovirus HBGAs gene expression CgFUT1 |
| url | https://www.mdpi.com/1467-3045/45/5/267 |
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