Genetic Code Expansion System for Tight Control of Gene Expression in <i>Bombyx mori</i> Cell Lines
Inducible gene expression systems are important tools for studying gene function and to control protein synthesis. With the completion of the detailed map of the silkworm (<i>Bombyx mori</i>) genome, the study of <i>Bombyx mori</i> has entered the post-genome era. While the f...
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| Format: | Article |
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MDPI AG
2021-12-01
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| Series: | Insects |
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| Online Access: | https://www.mdpi.com/2075-4450/12/12/1081 |
| _version_ | 1797503574266085376 |
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| author | Wei Lu Ruolin Wang Pan Wang Sanyuan Ma Qingyou Xia |
| author_facet | Wei Lu Ruolin Wang Pan Wang Sanyuan Ma Qingyou Xia |
| author_sort | Wei Lu |
| collection | DOAJ |
| description | Inducible gene expression systems are important tools for studying gene function and to control protein synthesis. With the completion of the detailed map of the silkworm (<i>Bombyx mori</i>) genome, the study of <i>Bombyx mori</i> has entered the post-genome era. While the functions of many genes have been described in detail, many coding genes remain unidentified. Except for the available tetracycline induction system, there is currently a dearth of other effective induction systems for <i>B. mori</i>. A genetic code expansion system can be used for protein labeling and to regulate gene expression. Here, we have established a genetic code expansion system for <i>B. mori</i> based on the well-researched tRNA<sup>Pyl</sup>/PylRS pair from <i>Methanosarcina mazei</i>. We used H-Lys(Boc)-OH, which is a lysine derivative to efficiently and tightly control the expression of the reporter gene DsRed[TAG]EGFP (D[TAG]G), which encoded a H-Lys(Boc)-OH-bearing protein fused with DsRed and EGFP (here regarded as D[Boc]G) in <i>B. mori</i> cell lines BmE and BmNs. In D[TAG]G, the amber stop codon is recognized as the orthogonal tRNA<sup>Pyl</sup>. Successful application of genetic code expansion system in silkworm cell lines will support the research into the function of silkworm genes and paves the way for the identification of new genes and protein markers in silkworm. |
| first_indexed | 2024-03-10T03:52:38Z |
| format | Article |
| id | doaj.art-5c76dd43d97643849484ec4e0965dd68 |
| institution | Directory Open Access Journal |
| issn | 2075-4450 |
| language | English |
| last_indexed | 2024-03-10T03:52:38Z |
| publishDate | 2021-12-01 |
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| record_format | Article |
| series | Insects |
| spelling | doaj.art-5c76dd43d97643849484ec4e0965dd682023-11-23T08:52:36ZengMDPI AGInsects2075-44502021-12-011212108110.3390/insects12121081Genetic Code Expansion System for Tight Control of Gene Expression in <i>Bombyx mori</i> Cell LinesWei Lu0Ruolin Wang1Pan Wang2Sanyuan Ma3Qingyou Xia4State Key Laboratory of Silkworm Genome Biology, Biological Science Research Center, Southwest University, Chongqing 400715, ChinaState Key Laboratory of Silkworm Genome Biology, Biological Science Research Center, Southwest University, Chongqing 400715, ChinaState Key Laboratory of Silkworm Genome Biology, Biological Science Research Center, Southwest University, Chongqing 400715, ChinaState Key Laboratory of Silkworm Genome Biology, Biological Science Research Center, Southwest University, Chongqing 400715, ChinaState Key Laboratory of Silkworm Genome Biology, Biological Science Research Center, Southwest University, Chongqing 400715, ChinaInducible gene expression systems are important tools for studying gene function and to control protein synthesis. With the completion of the detailed map of the silkworm (<i>Bombyx mori</i>) genome, the study of <i>Bombyx mori</i> has entered the post-genome era. While the functions of many genes have been described in detail, many coding genes remain unidentified. Except for the available tetracycline induction system, there is currently a dearth of other effective induction systems for <i>B. mori</i>. A genetic code expansion system can be used for protein labeling and to regulate gene expression. Here, we have established a genetic code expansion system for <i>B. mori</i> based on the well-researched tRNA<sup>Pyl</sup>/PylRS pair from <i>Methanosarcina mazei</i>. We used H-Lys(Boc)-OH, which is a lysine derivative to efficiently and tightly control the expression of the reporter gene DsRed[TAG]EGFP (D[TAG]G), which encoded a H-Lys(Boc)-OH-bearing protein fused with DsRed and EGFP (here regarded as D[Boc]G) in <i>B. mori</i> cell lines BmE and BmNs. In D[TAG]G, the amber stop codon is recognized as the orthogonal tRNA<sup>Pyl</sup>. Successful application of genetic code expansion system in silkworm cell lines will support the research into the function of silkworm genes and paves the way for the identification of new genes and protein markers in silkworm.https://www.mdpi.com/2075-4450/12/12/1081<i>Bombyx mori</i>gene expressiongenetic code expansion system |
| spellingShingle | Wei Lu Ruolin Wang Pan Wang Sanyuan Ma Qingyou Xia Genetic Code Expansion System for Tight Control of Gene Expression in <i>Bombyx mori</i> Cell Lines Insects <i>Bombyx mori</i> gene expression genetic code expansion system |
| title | Genetic Code Expansion System for Tight Control of Gene Expression in <i>Bombyx mori</i> Cell Lines |
| title_full | Genetic Code Expansion System for Tight Control of Gene Expression in <i>Bombyx mori</i> Cell Lines |
| title_fullStr | Genetic Code Expansion System for Tight Control of Gene Expression in <i>Bombyx mori</i> Cell Lines |
| title_full_unstemmed | Genetic Code Expansion System for Tight Control of Gene Expression in <i>Bombyx mori</i> Cell Lines |
| title_short | Genetic Code Expansion System for Tight Control of Gene Expression in <i>Bombyx mori</i> Cell Lines |
| title_sort | genetic code expansion system for tight control of gene expression in i bombyx mori i cell lines |
| topic | <i>Bombyx mori</i> gene expression genetic code expansion system |
| url | https://www.mdpi.com/2075-4450/12/12/1081 |
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