Related Experiment Video
Updated: Jan 4, 2026
06:28
Standardized Modular Assembly of Polycistronic Operons with Modular Cloning (MoClo) using the In-Cloning toolkit
Published on: September 2, 2025
573
MoChlo: A Versatile, Modular Cloning Toolbox for Chloroplast Biotechnology.
Alessandro Occhialini1,2, Agnieszka A Piatek3, Alexander C Pfotenhauer1,2
1Department of Food Science, University of Tennessee, Knoxville, Tennessee 37996.
Plant Physiology
|January 26, 2019
Summary
The MoChlo system enables rapid chloroplast engineering by providing a modular cloning kit. This tool accelerates the design of synthetic pathways for producing valuable compounds in plants.
Area of Science:
- Plant synthetic biology
- Chloroplast biotechnology
Background:
- Synthetic biology tools are advancing rapidly, particularly for chloroplast engineering.
- Golden Gate assembly facilitates the construction of large DNA molecules for genetic engineering.
Purpose of the Study:
- To develop and validate a modular chloroplast cloning system (MoChlo) for fast and flexible plant chloroplast engineering.
- To create a standardized kit of genetic parts for efficient synthetic biology applications in chloroplasts.
Main Methods:
- Developed the MoChlo system using a library of 128 standardized chloroplast-specific parts.
- Utilized Golden Gate assembly for constructing synthetic genetic elements into larger DNA constructs.
- Validated the system by assembling synthetic operons of varying sizes and assessing assembly efficiency.
Main Results:
- Successfully generated chloroplast transformation vectors with up to seven transcriptional units (∼10.6-kb synthetic operon).
- Demonstrated the MoChlo system's capability for fast and flexible chloroplast engineering.
- Made the MoChlo kit available through Addgene for researchers with limited resources.
Conclusions:
- The MoChlo kit is a valuable tool for accelerating progress in chloroplast biotechnology.
- Facilitates the design of heterologous metabolic pathways for plastid metabolic engineering.
- Enables efficient construction of complex genetic circuits in plant chloroplasts.
Related Concept Videos
02:19Export of Mitochondrial and Chloroplast Genes
4.1K
A eukaryotic cell can have up to three different types of genetic systems: nuclear, mitochondrial, and chloroplast. During evolution, organelles have exported many genes to the nucleus; this transfer is still ongoing in some plant species. Approximately 18% of the Arabidopsis thaliana nuclear genome is thought to be derived from the chloroplast’s cyanobacterial ancestor, and around 75% of the yeast genome derived from the mitochondria’s bacterial ancestor. This export has occurred...
4.1K
01:27Reproductive Cloning
32.4K
Reproductive cloning is the process of producing a genetically identical copy—a clone—of an entire organism. While clones can be produced by splitting an early embryo—similar to what happens naturally with identical twins—cloning of adult animals is usually done by a process called somatic cell nuclear transfer (SCNT).
Somatic Cell Nuclear Transfer
In SCNT, an egg cell is taken from an animal and its nucleus is removed, creating an enucleated egg. Then a somatic...
Somatic Cell Nuclear Transfer
In SCNT, an egg cell is taken from an animal and its nucleus is removed, creating an enucleated egg. Then a somatic...
32.4K
02:50Transgenic Plants
8.3K
Recombinant DNA technology called transgenesis is often used to add a foreign gene or remove a detrimental gene from an organism. Such genetically modified organisms are called transgenic organisms.
The first-ever transgenic plant was a tobacco plant developed in 1983 that showed resistance against the tobacco mosaic virus. Since then, many transgenic plants have been developed and commercialized for improving the agricultural, ornamental, and horticultural value of a crop plant. Transgenic...
The first-ever transgenic plant was a tobacco plant developed in 1983 that showed resistance against the tobacco mosaic virus. Since then, many transgenic plants have been developed and commercialized for improving the agricultural, ornamental, and horticultural value of a crop plant. Transgenic...
8.3K
01:08Cloning of Dolly the Sheep
6.9K
The first successfully cloned mammal was Dolly, a sheep, born on 5th July 1996 at Roslin Institute, Scotland. The cloned sheep was named after the American singer Dolly Parton. Dolly lived for seven years and died of respiratory complications, which is speculated to be due to the actual age of her DNA. Because the DNA in cloned cells belongs to an older individual, the cloned individual’s life expectancy may be affected. Indeed, analysis of Dolly’s DNA revealed shorter...
6.9K
01:59Plant Breeding and Biotechnology
21.3K
Crop cultivation has a long history in human civilization, with records showing the cultivation of cereal plants beginning at around 8000 BC. This early plant breeding was developed primarily to provide a steady supply of food.
21.3K