Stable expression of human tissue-type plasminogen activator regulated by beta-actin promoter in three human cell lines: HeLa, WI-38 VA13 and KMS-5.
A high-level and stable expression system of human tissue-type plasminogen activator (t-PA) was accomplished in human cells by selecting a promoter and a host cell line. First, we have constructed two types of t-PA expression plasmids containing 3 kb of the human beta-actin promoter region or 0.3 kb of SV40 early promoter region and these plasmids were transfected into HeLa cells, respectively, and the resulting transfectants were found to secrete various amounts of t-PA derived from the plasmids to the culture media. Southern blot analysis revealed that the beta-actin promoter was more efficient than the SV40 early promoter with regard to the expression level per single copy of the t-PA gene in the transfected HeLa cells. Next, the t-PA expression plasmid containing the beta-actin promoter was also transfected into WI-38 VA13 cells, a human fibroblastic cell line, and KMS-5 cells, a human lymphoid cell line, in order to compare the expression ability of the promoter among these three cell lines. Some of the transfectants from both cell lines were also found to produce t-PA. It was also found that the expression levels in HeLa and WI-38 VA13 seemed to be more efficient than that in KMS-5.
Biosciences Research Laboratory, Mochida Pharmaceutical Co., Tokyo, Japan.
H
H
Morishita
N
N
Nakamura
T
T
Yamakawa
H
H
Ogino
T
T
Kanamori
M
M
Nobuhara
M
M
Namba
D000199:Actins / Q000235:genetics*; D002460:Cell Line; D005347:Fibroblasts; D015870:Gene Expression; D006367:HeLa Cells; D006801:Humans; D010957:Plasmids; D011401:Promoter Regions, Genetic; D011994:Recombinant Proteins / Q000096:biosynthesis*; D010959:Tissue Plasminogen Activator / Q000096:biosynthesis* / Q000235:genetics; D014162:Transfection
D000199:Actins; D011994:Recombinant Proteins; D010959:Tissue Plasminogen Activator