Title |
A Modular Cloning System for Standardized Assembly of Multigene Constructs
|
---|---|
Published in |
PLOS ONE, February 2011
|
DOI | 10.1371/journal.pone.0016765 |
Pubmed ID | |
Authors |
Ernst Weber, Carola Engler, Ramona Gruetzner, Stefan Werner, Sylvestre Marillonnet |
Abstract |
The field of synthetic biology promises to revolutionize biotechnology through the design of organisms with novel phenotypes useful for medicine, agriculture and industry. However, a limiting factor is the ability of current methods to assemble complex DNA molecules encoding multiple genetic elements in various predefined arrangements. We present here a hierarchical modular cloning system that allows the creation at will and with high efficiency of any eukaryotic multigene construct, starting from libraries of defined and validated basic modules containing regulatory and coding sequences. This system is based on the ability of type IIS restriction enzymes to assemble multiple DNA fragments in a defined linear order. We constructed a 33 kb DNA molecule containing 11 transcription units made from 44 individual basic modules in only three successive cloning steps. This modular cloning (MoClo) system can be readily automated and will be extremely useful for applications such as gene stacking and metabolic engineering. |
X Demographics
Geographical breakdown
Country | Count | As % |
---|---|---|
Germany | 1 | 17% |
France | 1 | 17% |
United States | 1 | 17% |
United Kingdom | 1 | 17% |
Unknown | 2 | 33% |
Demographic breakdown
Type | Count | As % |
---|---|---|
Scientists | 4 | 67% |
Members of the public | 2 | 33% |
Mendeley readers
Geographical breakdown
Country | Count | As % |
---|---|---|
United States | 14 | <1% |
Germany | 12 | <1% |
United Kingdom | 8 | <1% |
Canada | 6 | <1% |
France | 3 | <1% |
Belgium | 3 | <1% |
Israel | 1 | <1% |
South Africa | 1 | <1% |
Sweden | 1 | <1% |
Other | 8 | <1% |
Unknown | 2103 | 97% |
Demographic breakdown
Readers by professional status | Count | As % |
---|---|---|
Student > Ph. D. Student | 458 | 21% |
Researcher | 349 | 16% |
Student > Bachelor | 329 | 15% |
Student > Master | 303 | 14% |
Student > Doctoral Student | 74 | 3% |
Other | 190 | 9% |
Unknown | 457 | 21% |
Readers by discipline | Count | As % |
---|---|---|
Agricultural and Biological Sciences | 761 | 35% |
Biochemistry, Genetics and Molecular Biology | 695 | 32% |
Engineering | 66 | 3% |
Chemistry | 33 | 2% |
Immunology and Microbiology | 20 | <1% |
Other | 103 | 5% |
Unknown | 482 | 22% |