scholarly journals Komagataeibacter Tool Kit (KTK): A Modular Cloning System for Multigene Constructs and Programmed Protein Secretion from Cellulose Producing Bacteria

2021 ◽  
Author(s):  
Vivianne J. Goosens ◽  
Kenneth T. Walker ◽  
Silvia M. Aragon ◽  
Amritpal Singh ◽  
Vivek R. Senthivel ◽  
...  
Keyword(s):  
Protein Secretion ◽  
Cloning System ◽  
Modular Cloning

scholarly journals Komagataeibacter tool kit (KTK): a modular cloning system for multigene constructs and programmed protein secretion from cellulose producing bacteria

2021 ◽  
Author(s):  
Vivianne J Goosens ◽  
Kenneth T Walker ◽  
Silvia M Aragon ◽  
Amritpal Singh ◽  
Vivek R Senthivel ◽  
...  
Keyword(s):  
Synthetic Biology ◽  
Protein Secretion ◽  
Functional Expression ◽  
Dna Assembly ◽  
Cloning System ◽  
Pure Cellulose ◽  
Modular Cloning ◽  
High Yields ◽  
Short Time ◽  
Cellulose Materials

Bacteria proficient at producing cellulose are an attractive synthetic biology host for the emerging field of Engineered Living Materials (ELMs). Species from the Komagataeibacter genus produce high yields of pure cellulose materials in a short time with minimal resources, and pioneering work has shown that genetic engineering in these strains is possible and can be used to modify the material and its production. To accelerate synthetic biology progress in these bacteria, we introduce here the Komagataeibacter tool kit (KTK), a standardised modular cloning system based on Golden Gate DNA assembly that allows DNA parts to be combined to build complex multigene constructs expressed in bacteria from plasmids. Working in Komagataeibacter rhaeticus, we describe basic parts for this system, including promoters, fusion tags and reporter proteins, before showcasing how the assembly system enables more complex designs. Specifically, we use KTK cloning to reformat the Escherichia coli curli amyloid fibre system for functional expression in K. rhaeticus, and go on to modify it as a system for programming protein secretion from the cellulose producing bacteria. With this toolkit, we aim to accelerate modular synthetic biology in these bacteria, and enable more rapid progress in the emerging ELMs community.

scholarly journals Peripheral infrastructure vectors and an extended set of plant parts for the modular cloning system

2017 ◽  
Author(s):  
Johannes Gantner ◽  
Theresa Ilse ◽  
Jana Ordon ◽  
Carola Kretschmer ◽  
Ramona Gruetzner ◽  
...  
Keyword(s):  
Building Blocks ◽  
Research Community ◽  
Dna Assembly ◽  
Golden Gate ◽  
Hierarchical Assembly ◽  
Cloning System ◽  
Plant Parts ◽  
Modular Cloning ◽  
Cost Efficient ◽  
Combinatorial Assembly

AbstractStandardized DNA assembly strategies facilitate the generation of multigene constructs from collections of building blocks in plant synthetic biology. A common syntax for hierarchical DNA assembly following the Golden Gate principle employing Type IIs restriction endonucleases was recently developed, and underlies the Modular Cloning and GoldenBraid systems. In these systems, transcriptional units and/or multigene constructs are assembled from libraries of standardized building blocks, also referred to as phytobricks, in several hierarchical levels and by iterative Golden Gate reactions. This combinatorial assembly strategy meets the increasingly complex demands in biotechnology and bioengineering, and also represents a cost-efficient and versatile alternative to previous molecular cloning techniques. For Modular Cloning, a collection of commonly used Plant Parts was previously released together with the Modular Cloning toolkit itself, which largely facilitated the adoption of this cloning system in the research community. Here, a collection of approximately 80 additional phytobricks is provided. These phytobricks comprise e.g. modules for inducible expression systems, different promoters or epitope tags, which will increase the versatility of Modular Cloning-based DNA assemblies. Furthermore, first instances of a “peripheral infrastructure” around Modular Cloning are presented: While available toolkits are designed for the assembly of plant transformation constructs, vectors were created to also use coding sequence-containing phytobricks directly in yeast two hybrid interaction or bacterial infection assays. Additionally, DNA modules and assembly strategies for connecting Modular Cloning with Gateway Cloning are presented, which may serve as an interface between available resources and newly adopted hierarchical assembly strategies. The presented material will be provided as a toolkit to the plant research community and will further enhance the usefulness and versatility of Modular Cloning.

scholarly journals GoldenPiCS: a Golden Gate-derived modular cloning system for applied synthetic biology in the yeast Pichia pastoris

2017 ◽  
Vol 11 (1) ◽  
Author(s):  
Roland Prielhofer ◽  
Juan J. Barrero ◽  
Stefanie Steuer ◽  
Thomas Gassler ◽  
Richard Zahrl ◽  
...  
Keyword(s):  
Synthetic Biology ◽  
Pichia Pastoris ◽  
Golden Gate ◽  
Yeast Pichia Pastoris ◽  
Cloning System ◽  
Modular Cloning

scholarly journals Adaptation of the GoldenBraid modular cloning system and creation of a toolkit for the expression of heterologous proteins in yeast mitochondria

2017 ◽  
Vol 17 (1) ◽  
Author(s):  
Ana Pérez-González ◽  
Ryan Kniewel ◽  
Marcel Veldhuizen ◽  
Hemant K. Verma ◽  
Mónica Navarro-Rodríguez ◽  
...  
Keyword(s):  
Yeast Mitochondria ◽  
Heterologous Proteins ◽  
Cloning System ◽  
Modular Cloning

scholarly journals A Modular Cloning System for Standardized Assembly of Multigene Constructs

PLoS ONE ◽  
2011 ◽  
Vol 6 (2) ◽  
pp. e16765 ◽  
Author(s):  
Ernst Weber ◽  
Carola Engler ◽  
Ramona Gruetzner ◽  
Stefan Werner ◽  
Sylvestre Marillonnet
Keyword(s):  
Cloning System ◽  
Modular Cloning

scholarly journals Peripheral infrastructure vectors and an extended set of plant parts for the Modular Cloning system

PLoS ONE ◽  
2018 ◽  
Vol 13 (5) ◽  
pp. e0197185 ◽  
Author(s):  
Johannes Gantner ◽  
Jana Ordon ◽  
Theresa Ilse ◽  
Carola Kretschmer ◽  
Ramona Gruetzner ◽  
...  
Keyword(s):  
Cloning System ◽  
Plant Parts ◽  
Modular Cloning
Sign in / Sign up

Export Citation Format

Share Document