The complete sequences of plasmids pFNeo and pMH-Neo: convenient expression vectors for high-level expression of eukaryotic genes in hematopoietic cell lines.
The eukaryotic expression vector, pFNeo, confers resistance to the antibiotic G418 and directs hematopoietic-specific expression of proteins under the control of the long terminal repeat from the Friend spleen focus-forming virus. Here, we report the entire nucleotide sequence of pFNeo. We have also constructed and sequenced a pFNeo-based expression vector (pMH-Neo) that carries an improved multiple cloning site region for easier subcloning. The utility of both vectors was demonstrated by transfection of murine T-cell hybridomas by electroporation. Transfection with either pFNeo or pMH-Neo yielded a high frequency (1 in 2 x 10(4)) of G418-resistant cell lines.
Division of Pediatric Oncology, Dana-Farber Cancer Institute, Boston, MA 02115.
W C
WC
Hahn
E
E
Menzin
T
T
Saito
R N
RN
Germain
B E
BE
Bierer
D000818:Animals; D000900:Anti-Bacterial Agents / Q000494:pharmacology; D001483:Base Sequence; D002460:Cell Line; D003001:Cloning, Molecular / Q000379:methods*; D005622:Friend murine leukemia virus / Q000235:genetics; D015870:Gene Expression; D005822:Genetic Vectors; D005839:Gentamicins / Q000494:pharmacology; D006410:Hematopoiesis; D006801:Humans; D008969:Molecular Sequence Data; D009838:Oligodeoxyribonucleotides; D010957:Plasmids; D012091:Repetitive Sequences, Nucleic Acid; D015183:Restriction Mapping; D013155:Spleen Focus-Forming Viruses / Q000235:genetics; D014162:Transfection / Q000379:methods*
D000900:Anti-Bacterial Agents; D005839:Gentamicins; D009838:Oligodeoxyribonucleotides; C010680:antibiotic G 418