New vectors for the efficient expression of mammalian genes in cultured cells.
We have constructed a new pair of plasmid vectors for the efficient expression of mammalian genes. The first of the new plasmids, pAVE1, was derived from pCMVcat [Foecking and Hofstetter, Gene 45 (1986) 101-105] by replacing the chloramphenicol acetyltransferase-encoding sequences in the latter for a multiple cloning site. Since it possesses the powerful enhancer-promoter unit of the immediate early gene of human cytomegalovirus, pAVE1 is ideal for the expression of mammalian genes. The second expression vector, pAVE2, resulted when the 3'-end flanking region from the human growth hormone-encoding gene (hGH) was incorporated in pAVE1. This region provides sequences for 3'-end processing and polyadenylation of primary transcripts. Thus, pAVE2 is suitable for expression of cDNAs in cultured cells, where introns have little effect on gene expression. To test our new vectors, we inserted the structural region of the chromosomal hGH gene into pAVE1, and its cDNA into pAVE2. By independently transfecting the resulting recombinant plasmids into COS-7 cells, we have achieved high levels of hGH transient expression with both vectors.
Ramírez-Solís
R
R
U.L.I.E.G. Departamento de Bioquimica, Facultad de Medicina, Monterrey, N.L., México.
Reséndez-Pérez
D
D
Alvidrez-Quihui
L E
LE
Rincón-Limas
D E
DE
Varela-Martínez
R
R
Martínez-Rodríguez
H G
HG
Barrera-Saldaña
H A
HA
D002478:Cells, Cultured; D015500:Chloramphenicol O-Acetyltransferase / Q000235:genetics / Q000378:metabolism; D003001:Cloning, Molecular; D003587:Cytomegalovirus / Q000235:genetics; D004742:Enhancer Elements, Genetic; D005786:Gene Expression Regulation; D005796:Genes; D005822:Genetic Vectors; D013006:Growth Hormone / Q000235:genetics; D006801:Humans; D010957:Plasmids; D011401:Promoter Regions, Genetic
D013006:Growth Hormone; D015500:Chloramphenicol O-Acetyltransferase