A novel expression vector composed of a regulatory element of the human leukosialin-encoding gene in different types of mammalian cells.
The regulatory element (RE) of the human leukosialin (LS)-encoding gene, that encodes a major sialoglycoprotein of human leukocyte and platelet membranes, was used to develop a novel expression vector, pKX. The vector was constructed by cloning a RE fragment and the SV40 fragment containing polyadenylation and splicing signals between HindIII and BamHI sites of the pCAT-Basic vector. The transcription level controlled by this vector was evaluated in six different cell lines using a transient expression assay of chloramphenicol acetyltransferase (CAT). The CAT activity of the pKX vector was compared to the other common expression vectors, namely pMSG (driven by the mouse mammary tumor virus LTR), pcDL-SR alpha (SV40 promoter/enhancer and HTLV-I LTR), pcDNAI (cytomegalovirus promoter/enhancer) and pCAT-Control (SV40 promoter/enhancer). The level of expression provided by the pKX vector was comparable to that observed with pcDNAI and pcDL-SR alpha vectors. In different mammalian cell lines, the highest efficiency of expression of the pKX vector was observed in the human T-cell lines, Jurkat and CEM, although the expression of pcDL-SR alpha-CAT in those cell lines was in the same range. The expression of the pKX vector driven by a non-viral promoter and/or enhancer can be as efficient as that driven by a viral promoter and/or enhancer. Potential uses of this vector may be found in studies of transient gene expression in hematopoietic cells and for gene therapy, particularly the ones involving T-cells.
Xu
A
A
Alliance Pharmaceutical Corp., San Diego, CA 92121, USA.
Kudo
S
S
Fukuda
M
M
D000818:Animals; D015703:Antigens, CD; D001483:Base Sequence; D002478:Cells, Cultured; D015500:Chloramphenicol O-Acetyltransferase / Q000235:genetics; D003001:Cloning, Molecular / Q000379:methods*; D003587:Cytomegalovirus / Q000235:genetics; D004742:Enhancer Elements, Genetic; D017930:Genes, Reporter; D015316:Genetic Therapy; D005822:Genetic Vectors / Q000235:genetics*; D015368:Human T-lymphotropic virus 1 / Q000235:genetics; D006801:Humans; D015459:Leukemia-Lymphoma, Adult T-Cell / Q000473:pathology; D051917:Leukosialin; D008322:Mammals; D008324:Mammary Tumor Virus, Mouse / Q000235:genetics; D008969:Molecular Sequence Data; D009928:Organ Specificity; D011401:Promoter Regions, Genetic / Q000235:genetics*; D012045:Regulatory Sequences, Nucleic Acid; D012091:Repetitive Sequences, Nucleic Acid; D012795:Sialoglycoproteins / Q000235:genetics*; D013539:Simian virus 40 / Q000235:genetics; D013601:T-Lymphocytes / Q000378:metabolism; D014158:Transcription, Genetic; D014407:Tumor Cells, Cultured
D015703:Antigens, CD; D051917:Leukosialin; C465014:SPN protein, human; D012795:Sialoglycoproteins; D015500:Chloramphenicol O-Acetyltransferase