Related Experiment Video
Updated: Jun 13, 2026
14:07
Genome-wide Gene Deletions in Streptococcus sanguinis by High Throughput PCR
Published on: November 23, 2012
Two pHZ1358-derivative vectors for efficient gene knockout in streptomyces
Yunlong He1, Zhijun Wang, Linquan Bai
1Laboratory of Microbiol Metabolism, Shanghai Jiaotong University, Shanghai. China.
Journal of Microbiology and Biotechnology
|May 15, 2010
Summary
Researchers optimized Streptomyces gene editing vectors for easier gene disruption and replacement. New plasmids, pJTU1278 and pJTU1289, enhance cloning and PCR-targeting in Streptomyces avermitilis, simplifying genetic manipulation.
Area of Science:
- Molecular Biology
- Microbial Genetics
- Synthetic Biology
Background:
- Streptomyces plasmids like pIJ101 are crucial for genetic manipulation.
- Previous derivatives like pHZ1358 enabled gene disruption and replacement.
- Further optimization was needed for enhanced cloning and targeted gene editing.
Purpose of the Study:
- To engineer improved Streptomyces vectors for gene editing.
- To facilitate plasmid construction in E. coli and enable PCR-targeting.
- To demonstrate the utility of the new vectors in S. avermitilis.
Main Methods:
- Construction of a new plasmid derivative, pJTU1278, incorporating multiple cloning sites and a lacZ marker.
- Deletion of the oriT region from pJTU1278 to create pJTU1289 for PCR-targeting.
- Application of the developed vectors for gene deletion in Streptomyces avermitilis.
Main Results:
- pJTU1278 facilitates convenient plasmid construction in E. coli.
- pJTU1289 serves as a specialized vector for PCR-targeting applications.
- Successful deletion of an avermectin biosynthetic gene in S. avermitilis using the new vectors.
Conclusions:
- The optimized vectors pJTU1278 and pJTU1289 represent significant advancements for Streptomyces genetic engineering.
- These tools streamline gene disruption, replacement, and targeted gene editing.
- The study validates the enhanced efficiency and applicability of these novel vectors.
Related Concept Videos
01:16In-vitro Mutagenesis
To learn more about the function of a gene, researchers can observe what happens when the gene is inactivated or “knocked out,” by creating genetically engineered knockout animals. Knockout mice have been particularly useful as models for human diseases such as cancer, Parkinson’s disease, and diabetes.
02:53Conservative 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...
The recognition sites for Cre recombinase called LoxP...