title: One-step construction of lentiviral reporter using Red-mediated recombination.
abstract: Current approaches to generate lentiviral vectors, which have been used extensively for gene therapy, are time consuming and require a large expenditure. Here, we directly clone the full length myosin light chain kinase cDNA into enhanced green fluorescence protein (EGFP)-fused pLenti6/V5 expression vector in just one step with the use of Red-mediated recombination system, allowing for rapid and effective cloning of lentiviral expression vectors. In addition, the simultaneous expression of EGFP reporter provides a convenient monitoring mean for host cell infection and for localization of the target proteins.
authors.affiliation: Department of Geratology, The First Hospital Affiliated to Nanjing Medical University, Nanjing 210029, China.
authors.forename: Juanmin
authors.initials: J
authors.lastname: Zha
authors.forename: Xin
authors.initials: X
authors.lastname: Chen
authors.forename: Chaojun
authors.initials: C
authors.lastname: Li
authors.forename: Minsheng
authors.initials: M
authors.lastname: Zhu
authors.forename: Guoxian
authors.initials: G
authors.lastname: Ding
authors.forename: Weiqi
authors.initials: W
authors.lastname: He
mesh_terms: D001709:Biotechnology / Q000379:methods*; D005822:Genetic Vectors / Q000235:genetics*; D049452:Green Fluorescent Proteins / Q000378:metabolism; D057809:HEK293 Cells; D006801:Humans; D016086:Lentivirus / Q000235:genetics*; D009219:Myosin-Light-Chain Kinase / Q000378:metabolism; D010957:Plasmids / Q000235:genetics; D011995:Recombination, Genetic / Q000235:genetics*; D014162:Transfection
chemical_list: C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins; D009219:Myosin-Light-Chain Kinase