Overlapping promoter library designed for rational heterogenous expression in Cordyceps militaris
Abstract Background Cordyceps militaris, a kind of edible and medicinal fungus widely accepted in East Asia, has attracted much attention as a potential cell factory for producing adenosine analogs. Despite the rapid development in gene editing techniques and genome modeling, the diversity of DNA el...
| Main Authors: | , , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
BMC
2022-06-01
|
| Series: | Microbial Cell Factories |
| Subjects: | |
| Online Access: | https://doi.org/10.1186/s12934-022-01826-0 |
| _version_ | 1811257920309952512 |
|---|---|
| author | Mengdi Lyu Jiapeng Zeng Yue Zhou Tongyu Zhang Aiping Wang Jiezhao Ma Ziyi Wu Alvaro Castells-Garcia Esther González-Almela Junfang Lin Tao Wei |
| author_facet | Mengdi Lyu Jiapeng Zeng Yue Zhou Tongyu Zhang Aiping Wang Jiezhao Ma Ziyi Wu Alvaro Castells-Garcia Esther González-Almela Junfang Lin Tao Wei |
| author_sort | Mengdi Lyu |
| collection | DOAJ |
| description | Abstract Background Cordyceps militaris, a kind of edible and medicinal fungus widely accepted in East Asia, has attracted much attention as a potential cell factory for producing adenosine analogs. Despite the rapid development in gene editing techniques and genome modeling, the diversity of DNA elements in C. militaris was too short to achieve rational heterogeneous expression for metabolic engineering studies. Results In this study, PtrpC, a kind of promoter with a relatively appropriate expression level and small size, was selected as a monomer for promoter library construction. Through in vitro BioBricks assembly, 9 overlapping PtrpC promoters with different copy numbers as well as reporter gene gfp were connected and subsequently integrated into the genome of C. militaris. Both the mRNA transcription level and the expression level of gene gfp gradually increased along with the copy number of the overlapping promoter NPtrpC and peaked at 7. In the meantime, no significant difference was found in either the biomass or morphological characteristic of engineered and wild-type strains. Conclusions This study firstly expanded the overlapping promoter strategy used in model microorganism in C. militaris. It was a proof-of-concept in fungi synthetic biology and provide a general method to pushed the boundary of promoter engineering in edible mushroom. |
| first_indexed | 2024-04-12T18:04:56Z |
| format | Article |
| id | doaj.art-3540c105274f40d28bf8dc6ede0117de |
| institution | Directory Open Access Journal |
| issn | 1475-2859 |
| language | English |
| last_indexed | 2024-04-12T18:04:56Z |
| publishDate | 2022-06-01 |
| publisher | BMC |
| record_format | Article |
| series | Microbial Cell Factories |
| spelling | doaj.art-3540c105274f40d28bf8dc6ede0117de2022-12-22T03:22:00ZengBMCMicrobial Cell Factories1475-28592022-06-0121111010.1186/s12934-022-01826-0Overlapping promoter library designed for rational heterogenous expression in Cordyceps militarisMengdi Lyu0Jiapeng Zeng1Yue Zhou2Tongyu Zhang3Aiping Wang4Jiezhao Ma5Ziyi Wu6Alvaro Castells-Garcia7Esther González-Almela8Junfang Lin9Tao Wei10Department of Bioengineering, College of Food Science, South China Agricultural UniversityDepartment of Bioengineering, College of Food Science, South China Agricultural UniversityDepartment of Bioengineering, College of Food Science, South China Agricultural UniversityDepartment of Bioengineering, College of Food Science, South China Agricultural UniversityBioland LaboratoryDepartment of Bioengineering, College of Food Science, South China Agricultural UniversityDepartment of Bioengineering, College of Food Science, South China Agricultural UniversityBioland LaboratoryBioland LaboratoryDepartment of Bioengineering, College of Food Science, South China Agricultural UniversityDepartment of Bioengineering, College of Food Science, South China Agricultural UniversityAbstract Background Cordyceps militaris, a kind of edible and medicinal fungus widely accepted in East Asia, has attracted much attention as a potential cell factory for producing adenosine analogs. Despite the rapid development in gene editing techniques and genome modeling, the diversity of DNA elements in C. militaris was too short to achieve rational heterogeneous expression for metabolic engineering studies. Results In this study, PtrpC, a kind of promoter with a relatively appropriate expression level and small size, was selected as a monomer for promoter library construction. Through in vitro BioBricks assembly, 9 overlapping PtrpC promoters with different copy numbers as well as reporter gene gfp were connected and subsequently integrated into the genome of C. militaris. Both the mRNA transcription level and the expression level of gene gfp gradually increased along with the copy number of the overlapping promoter NPtrpC and peaked at 7. In the meantime, no significant difference was found in either the biomass or morphological characteristic of engineered and wild-type strains. Conclusions This study firstly expanded the overlapping promoter strategy used in model microorganism in C. militaris. It was a proof-of-concept in fungi synthetic biology and provide a general method to pushed the boundary of promoter engineering in edible mushroom.https://doi.org/10.1186/s12934-022-01826-0Cordyceps militarisFungiOverlapping promoterBioBricks |
| spellingShingle | Mengdi Lyu Jiapeng Zeng Yue Zhou Tongyu Zhang Aiping Wang Jiezhao Ma Ziyi Wu Alvaro Castells-Garcia Esther González-Almela Junfang Lin Tao Wei Overlapping promoter library designed for rational heterogenous expression in Cordyceps militaris Microbial Cell Factories Cordyceps militaris Fungi Overlapping promoter BioBricks |
| title | Overlapping promoter library designed for rational heterogenous expression in Cordyceps militaris |
| title_full | Overlapping promoter library designed for rational heterogenous expression in Cordyceps militaris |
| title_fullStr | Overlapping promoter library designed for rational heterogenous expression in Cordyceps militaris |
| title_full_unstemmed | Overlapping promoter library designed for rational heterogenous expression in Cordyceps militaris |
| title_short | Overlapping promoter library designed for rational heterogenous expression in Cordyceps militaris |
| title_sort | overlapping promoter library designed for rational heterogenous expression in cordyceps militaris |
| topic | Cordyceps militaris Fungi Overlapping promoter BioBricks |
| url | https://doi.org/10.1186/s12934-022-01826-0 |
| work_keys_str_mv | AT mengdilyu overlappingpromoterlibrarydesignedforrationalheterogenousexpressionincordycepsmilitaris AT jiapengzeng overlappingpromoterlibrarydesignedforrationalheterogenousexpressionincordycepsmilitaris AT yuezhou overlappingpromoterlibrarydesignedforrationalheterogenousexpressionincordycepsmilitaris AT tongyuzhang overlappingpromoterlibrarydesignedforrationalheterogenousexpressionincordycepsmilitaris AT aipingwang overlappingpromoterlibrarydesignedforrationalheterogenousexpressionincordycepsmilitaris AT jiezhaoma overlappingpromoterlibrarydesignedforrationalheterogenousexpressionincordycepsmilitaris AT ziyiwu overlappingpromoterlibrarydesignedforrationalheterogenousexpressionincordycepsmilitaris AT alvarocastellsgarcia overlappingpromoterlibrarydesignedforrationalheterogenousexpressionincordycepsmilitaris AT esthergonzalezalmela overlappingpromoterlibrarydesignedforrationalheterogenousexpressionincordycepsmilitaris AT junfanglin overlappingpromoterlibrarydesignedforrationalheterogenousexpressionincordycepsmilitaris AT taowei overlappingpromoterlibrarydesignedforrationalheterogenousexpressionincordycepsmilitaris |