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Poly-ε-caprolactone (PCL) incorporated with plasmid DNA was electrospun, and the resultant nanofibers were used to observe DNA release from the nanofibers. The plasmid DNA enhanced green fluorescent protein (EGFP) with cytomegalovirus (CMV) promoter (PCMVb-GFP) was amplified with E. coli. PCL was chosen because it is biodegradable aliphatic polyester, which plays a critical role in tissue engineering, such as scaffolding, drug, DNA, gene and protein delivery vehicles. Some of the physical and biological properties of the nanofibers were determined using different methods. Scanning electron microscopy (SEM) micrographs showed that nanofibers have an average diameter of about 100 nm. Cytotoxicity tests showed that cell viability for 1 day, 4 days, and 7 days of the tests were above 80 %. These data demonstrated that PCL-plasmid DNA nanofibers have no cytotoxicity and showed benign biocompatibility for biomedical applications. PCMVb-GFP plasmid-linked electrospun nanofibers continuously released double-stranded DNA for at least seven days. For the first 15 min, there was a burst release of about 1.8 ng/ml. For the following hours and days, the release was about to be the same (release of 0.575 ng/ml). Therefore, PCL nanofibers may be an ideal candidate for various biomedical applications such as cancer treatment, scaffolding, tissue engineering, and protein delivery vehicles.