Related Experiment Video
Updated: Apr 1, 2026
11:04
Author Spotlight: Methods for Electroporation and Transformation Confirmation in Limosilactobacillus reuteri DSM20016
Published on: June 23, 2023
5.3K
Prophage recombinases-mediated genome engineering in Lactobacillus plantarum
Peng Yang1, Jing Wang2, Qingsheng Qi3
1State Key Laboratory of Microbial Technology, Shandong University, Jinan, 250100, People's Republic of China. fwjt63298@126.com.
Microbial Cell Factories
|October 7, 2015
Summary
Researchers developed a new recombineering system for Lactobacillus plantarum, enabling efficient, marker-free gene deletion. This advance simplifies genetic engineering in this important food-grade bacterium.
Area of Science:
- Microbiology
- Molecular Biology
- Genetic Engineering
Background:
- Lactobacillus plantarum is a key food-grade lactic acid bacterium (LAB) with industrial and medical applications.
- Conventional gene deletion methods in L. plantarum are inefficient and labor-intensive.
- Recombineering offers a powerful in vivo DNA engineering approach.
Purpose of the Study:
- To establish a double-stranded DNA (dsDNA) recombineering system in Lactobacillus plantarum.
- To develop a method for marker-free genetic manipulation of the L. plantarum chromosome.
Main Methods:
- Identification of key proteins (lp_0640, lp_0641, lp_0642) involved in dsDNA recombination from a prophage locus.
- Utilizing these proteins to mediate homologous recombination between dsDNA and genomic DNA.
- Development of a marker-free chromosomal manipulation technique.
Main Results:
- A functional dsDNA recombineering system was successfully established in L. plantarum.
- The system enables homologous recombination for genetic modification.
- A novel method for marker-free genetic manipulation of the L. plantarum chromosome was created.
Conclusions:
- The Lp_0640-41-42-mediated recombination system facilitates easy mutant screening.
- This method provides an alternative to existing genetic manipulation techniques.
- The system is expected to aid in understanding LAB probiotic functions and physiology.
Related Concept Videos
02:53Conservative Site-specific Recombination and Phase Variation
7.4K
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...
The recognition sites for Cre recombinase called LoxP...
7.4K
02:50Transgenic Plants
9.2K
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...
9.2K
01:09Recombinant DNA
104.8K
Overview
104.8K
01:20The Central Dogma
35.3K
The central dogma explains the flow of genetic information from DNA nucleotides to the amino acid sequence of proteins.
RNA is the Missing Link Between DNA and Proteins
In the early 1900s, scientists discovered that DNA stores all the information needed for cellular functions and that proteins perform most of these functions. However, the mechanisms of converting genetic information into functional proteins remained unknown for many years. Initially, it was believed that a single gene is...
RNA is the Missing Link Between DNA and Proteins
In the early 1900s, scientists discovered that DNA stores all the information needed for cellular functions and that proteins perform most of these functions. However, the mechanisms of converting genetic information into functional proteins remained unknown for many years. Initially, it was believed that a single gene is...
35.3K
01:26Transformation
1.3K
Microbial communities are dynamic environments where cell lysis releases free DNA into the surroundings. Other cells can take up this extracellular DNA through a process known as transformation.When a cell incorporates this foreign DNA into its genome, resulting in genetic modification, the process is known as transformation. Cells capable of this process are termed competent. Competence can be natural, as observed in certain bacteria and archaea, or artificially induced in the...
1.3K
01:16Viral Replication: Lysogenic Cycle
2.4K
The lysogenic cycle is a crucial viral replication strategy that allows bacteriophages to persist within host cells without immediately destroying them. This process is primarily observed in temperate phages, such as bacteriophage lambda (λ), which infects Escherichia coli. The cycle allows the viral genome to persist across bacterial generations while keeping host cells viable.Integration of the Viral GenomeUpon infection, bacteriophage lambda attaches to the bacterial surface and injects...
2.4K