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Related Experiment Video

Updated: Jan 15, 2026

CAPRRESI: Chimera Assembly by Plasmid Recovery and Restriction Enzyme Site Insertion

CAPRRESI: Chimera Assembly by Plasmid Recovery and Restriction Enzyme Site Insertion

Published on: June 25, 2017

SHARK: A Specialized Host for Assembling R6K Plasmids.

Shivang Hina-Nilesh Joshi¹, Christopher Jenkins², David Ulaeto²

¹School of Biological Sciences, University of Bristol, 24 Tyndall Avenue, Bristol BS8 1TQ, U.K.

ACS Synthetic Biology

|January 13, 2026

Summary

This summary is machine-generated.

Researchers developed new Pir E. coli strains, called SHARK, for efficient R6K plasmid maintenance and cloning. These strains improve cloning efficiency for challenging genome engineering projects, making R6K plasmid use more accessible.

Keywords:

Pir strains R6K plasmids cloning conditional replication genome integration lambda-RED

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Last Updated: Jan 15, 2026

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Published on: February 5, 2021

Area of Science:

Molecular Biology
Microbiology

Background:

R6K plasmids are vital for genome engineering but require specific Pir E. coli strains for maintenance.
Existing Pir strains often have limitations, including obscure genetic backgrounds and poor cloning efficiency.

Purpose of the Study:

To develop novel E. coli strains optimized for stable R6K plasmid maintenance and efficient cloning.
To create a resource for researchers undertaking complex genome engineering projects.

Main Methods:

Construction of SHARK strains derived from DH10B, incorporating a genome-encoded pir gene for R6K plasmid replication.
Integration of a λCI gene for tight, unconditional repression of specific plasmid-borne genes.
Characterization of SHARK strain performance in transformation efficiency with large and complex cloning reactions.

Main Results:

SHARK strains demonstrate >100-fold higher transformation efficiency compared to a commercial Pir strain.
The new strains facilitate stable maintenance of R6K plasmids.
SHARK strains are effective for cloning large and complex genetic constructs.

Conclusions:

SHARK strains represent a significant advancement for R6K plasmid cloning in genome engineering.
These strains address limitations of existing Pir strains, enhancing cloning efficiency and accessibility.
The public availability of SHARK strains and associated tools will benefit the research community.