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Expression and amplification of cloned rat liver tyrosine aminotransferase in nonhepatic cells

✍ Scribed by Kulwant K. Kohli; Robert H. Stellwagen

Publisher
John Wiley and Sons
Year
1990
Tongue
English
Weight
721 KB
Volume
142
Category
Article
ISSN
0021-9541

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✦ Synopsis

A full-length cDNA tor the rat liver enzyme tyrosine aminotransferase has been used to construct manimalian expression vectors by recombindnt DNA techniques. These vectors, which have employed either a simian viru5 40 or a Rous sarcoma virus promoter, were transfected into a variety of nonhepatic mammalian cell lines in culture. Transient expression of tyrosine aminotransferase was readily observed after transfection into monkey COS cells and mouse L cells. Stable clones that express cloned tyrosine aminotransferase have been isolated from mouse 1 cells, hamster Wgl a fibroblasts, and Chinese hamster ovary (CHO) cells. A vector capable of expressing both tyrosine aminotransferase and dihydrofolate reductase was stimulated to undergo amplification by treatment with methotrexate in a CHO cell line deficient in the latter enzyme. Levels of tyrosine aminotransferase as much as 50-fold higher than typically seen in glucocorticoidinduced hepatoma cells were achiewd in some CHO clones by this technique.

The tyrosine aminotransferase produced a t these highly amplified levels appeared structurally normal and had no major harmful effects on the cells.

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