Search research articles

ABOUT JoVE

Overview Leadership Blog JoVE Help Center

AUTHORS

Publishing Process Editorial Board Scope & Policies Peer Review FAQ Submit

LIBRARIANS

Testimonials Subscriptions Access Resources Library Advisory Board FAQ

RESEARCH

JoVE Journal Methods Collections JoVE Encyclopedia of Experiments Archive

EDUCATION

JoVE Core JoVE Business JoVE Science Education JoVE Lab Manual Faculty Resource Center Faculty Site

Terms & Conditions of Use

Related Concept Videos

Conservative Site-specific Recombination and Phase Variation

Conservative Site-specific Recombination and Phase Variation

Because the DNA segments are cut and reorganized in a direction-specific manner, site-specific recombination has emerged as an efficient genetic engineering technique. Flippase and Cyclization recombinases or Flp and Cre, respectively, are two members of the tyrosine recombinase family derived from bacteriophages, that are used to mediate site-specific DNA insertions, deletions, and targeted expression of proteins in mammalian cell lines.
The recognition sites for Cre recombinase called LoxP...

Homologous Recombination

Homologous Recombination

The basic reaction of homologous recombination (HR) involves two chromatids that contain DNA sequences sharing a significant stretch of identity. One of these sequences uses a strand from another as a template to synthesize DNA in an enzyme-catalyzed reaction. The final product is a novel amalgamation of the two substrates. To ensure an accurate recombination of sequences, HR is restricted to the S and G2 phases of the cell cycle. At these stages, the DNA has been replicated already and the...

Viral Recombination

Viral Recombination

Cells are sometimes infected by more than one virus at once. When two viruses disassemble to expose their genomes for replication in the same cell, similar regions of their genomes can pair together and exchange sequences in a process called recombination. Alternatively, viruses with segmented genomes can swap segments in a process called reassortment.

Recombinant DNA

Recombinant DNA

Recombinant DNA

Recombinant DNA

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by

Same author

Alternatively spliced killer-protector system confers S19-mediated hybrid male sterility in rice.

Nature communications·2026

Same author

The pectate lyase OsPSE1 mediates root cell-wall pectin turnover to regulate senescence in rice.

Plant physiology and biochemistry : PPB·2026

Same author

Plant base editing: a decade of progress and future applications.

aBIOTECH·2026

Same author

Engineering the efficient Exo-Cas12i2 for MITE manipulation in rice.

Journal of integrative plant biology·2026

Same author

TranC: an evolutionary bridge from transposons to CRISPR tools.

Science bulletin·2026

Same author

A tripartite pollen killer-protector system confers temperature-sensitive inter-subspecific reproductive isolation in rice.

Molecular plant·2025

Search research articles

Related Experiment Video

Updated: Apr 27, 2026

Subcloning Plus Insertion SPI - A Novel Recombineering Method for the Rapid Construction of Gene Targeting Vectors

Subcloning Plus Insertion SPI - A Novel Recombineering Method for the Rapid Construction of Gene Targeting Vectors

Published on: January 8, 2015

Robust multi-type plasmid modifications based on isothermal in vitro recombination.

Qin-Long Zhu¹, Zhong-Fang Yang¹, Qun-Yu Zhang¹

¹State Key Laboratory for Conservation and Utilization of Subtropical Agro-bioresources, College of Life Sciences, South China Agricultural University, Guangzhou 510642, China.

|July 11, 2014

Summary

This summary is machine-generated.

A new method uses isothermal in vitro recombination for seamless, multi-type plasmid modifications. This versatile molecular biology technique allows simultaneous gene insertion, deletion, and replacement in plasmids.

Keywords:

Isothermal in vitro recombination Plasmid modification

More Related Videos

Use of In Vivo Assembly for High-efficiency Plasmid Construction

Use of In Vivo Assembly for High-efficiency Plasmid Construction

Published on: February 7, 2025

In Vitro Directed Evolution of a Restriction Endonuclease with More Stringent Specificity

In Vitro Directed Evolution of a Restriction Endonuclease with More Stringent Specificity

Published on: March 25, 2020

Related Experiment Videos

Last Updated: Apr 27, 2026

Subcloning Plus Insertion SPI - A Novel Recombineering Method for the Rapid Construction of Gene Targeting Vectors

Subcloning Plus Insertion SPI - A Novel Recombineering Method for the Rapid Construction of Gene Targeting Vectors

Published on: January 8, 2015

Use of In Vivo Assembly for High-efficiency Plasmid Construction

Use of In Vivo Assembly for High-efficiency Plasmid Construction

Published on: February 7, 2025

In Vitro Directed Evolution of a Restriction Endonuclease with More Stringent Specificity

In Vitro Directed Evolution of a Restriction Endonuclease with More Stringent Specificity

Published on: March 25, 2020

Area of Science:

Molecular Biology
Synthetic Biology
Genetic Engineering

Background:

Efficient and versatile plasmid modification is crucial for molecular biology research.
Existing methods often lack the flexibility for multiple, simultaneous genetic alterations.
Seamless integration of diverse genetic elements into plasmids presents a challenge.

Purpose of the Study:

To develop a robust strategy for multi-type plasmid modifications.
To demonstrate the efficiency of isothermal in vitro recombination for complex plasmid engineering.
To provide a versatile tool for diverse applications in molecular biology.

Main Methods:

Utilized isothermal in vitro recombination technology for plasmid engineering.
Developed a strategy enabling modifications at any desired position within plasmids.
Achieved seamless integration of sequence modifications without scar sequences.

Main Results:

Successfully demonstrated simultaneous multi-type plasmid modifications.
Exemplified the method by inserting a GFP gene, deleting a 623-bp fragment, and replacing an ampicillin resistance gene with a kanamycin resistance gene.
Confirmed the efficiency and seamless nature of the recombination-based approach.

Conclusions:

The isothermal in vitro recombination-based strategy offers a powerful approach for multi-type plasmid modification.
This method provides flexibility for complex genetic engineering tasks.
The technique holds broad application prospects in molecular biology studies and synthetic biology.