title: piggyBac-mediated generation of stable transfectants with surface human leukocyte antigen expression from a small number of cells.
abstract: Techniques useful for isolating a series of stable transfectants from a small number of cells are important for high-throughput functional analyses of genes of interest (GOIs). This study employed piggyBac transposon vectors carrying an IRES-eGFP cassette to isolate stable transfectants. The 14 transfections performed produced stable transfectants; a small number of cells (5.7×10(4)) was sufficient even when liposomes were used to transfect non-adherent cells. Of note, there was a close relationship between eGFP fluorescence intensity and GOI (human leukocyte antigen) expression level. Therefore, the current system would be useful for the high-throughput acquisition of clones showing high GOI expression.
authors: Department of Molecular Life Science, Division of Basic Medical Science and Molecular Medicine, Tokai University School of Medicine, Isehara, Kanagawa, Japan.
authors: Hiromi
authors: H
authors: Miura
authors: Hidetoshi
authors: H
authors: Inoko
authors: Itsuro
authors: I
authors: Inoue
authors: Yoshinori
authors: Y
authors: Okada
authors: Masafumi
authors: M
authors: Tanaka
authors: Masahiro
authors: M
authors: Sato
authors: Masato
authors: M
authors: Ohtsuka
mesh_terms: D002452:Cell Count; D002460:Cell Line; D004251:DNA Transposable Elements / Q000235:genetics*; D015870:Gene Expression; D005822:Genetic Vectors / Q000235:genetics; D006680:HLA Antigens / Q000235:genetics*; D006801:Humans; D014162:Transfection / Q000379:methods*
chemical_list: D004251:DNA Transposable Elements; D006680:HLA Antigens