title: Robust multi-type plasmid modifications based on isothermal in vitro recombination.
abstract: A robust strategy for multi-type plasmid modifications is developed based on the isothermal in vitro recombination technology, by which any combination of the sequence modifications can be efficiently achieved in plasmids at any desired position in a seamless manner. As an example, we showed that a plasmid modification with insertion of a GFP gene, deletion of a 623-bp fragment, and substitution of an ampicillin resistance gene by a kanamycin resistance gene was accomplished simultaneously by this method. Therefore, the isothermal in vitro recombination-based multi-type plasmid modification strategy is a useful approach for broad application prospects in molecular biology studies.
authors.affiliation: State Key Laboratory for Conservation and Utilization of Subtropical Agro-bioresources, College of Life Sciences, South China Agricultural University, Guangzhou 510642, China. Electronic address: zhuql@scau.edu.cn.
authors.forename: Qin-Long
authors.initials: QL
authors.lastname: Zhu
authors.affiliation: State Key Laboratory for Conservation and Utilization of Subtropical Agro-bioresources, College of Life Sciences, South China Agricultural University, Guangzhou 510642, China.
authors.forename: Zhong-Fang
authors.initials: ZF
authors.lastname: Yang
authors.affiliation: State Key Laboratory for Conservation and Utilization of Subtropical Agro-bioresources, College of Life Sciences, South China Agricultural University, Guangzhou 510642, China.
authors.forename: Qun-Yu
authors.initials: QY
authors.lastname: Zhang
authors.affiliation: State Key Laboratory for Conservation and Utilization of Subtropical Agro-bioresources, College of Life Sciences, South China Agricultural University, Guangzhou 510642, China.
authors.forename: Le-Tian
authors.initials: LT
authors.lastname: Chen
authors.affiliation: State Key Laboratory for Conservation and Utilization of Subtropical Agro-bioresources, College of Life Sciences, South China Agricultural University, Guangzhou 510642, China. Electronic address: ygliu@scau.edu.cn.
authors.forename: Yao-Guang
authors.initials: YG
authors.lastname: Liu
mesh_terms: D000668:Ampicillin Resistance / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D005821:Genetic Techniques; D049452:Green Fluorescent Proteins / Q000235:genetics; D007613:Kanamycin Resistance / Q000235:genetics; D010957:Plasmids / Q000235:genetics*; D016133:Polymerase Chain Reaction; D011995:Recombination, Genetic; D017384:Sequence Deletion
chemical_list: D049452:Green Fluorescent Proteins
keywords: Isothermal in vitro recombination; Plasmid modification