Can I read Co-transformation of Lec1 CHO cells with N-acetylgucosaminyltransferase 1 activity and a selectable marker on EtoBox?

Co-transformation of Lec1 CHO cells with N-acetylgucosaminyltransferase 1 activity and a selectable marker by James Ripka; Michael Pierce; Nevis Fregien is a Biochemistry, Genetics and Molecular Biology article available to read on EtoBox.

What is Co-transformation of Lec1 CHO cells with N-acetylgucosaminyltransferase 1 activity and a selectable marker about?

In animal cells, the enzyme a( 1,3)-mannoside-P( 1,2)-N-acetylglucosaminyltransferase I (GlcNAc-TI, EC.2.4.1.101) catalyzes the addition of N-acetylglucosamine to the ASN-linked Man GlcNAc oligosaccharide. The Chinese hamster ovary (CHO) mutant cell line Lecl is deficient in this enzyme activity and, therefore, accumulates mannose-terminating cell surface ASN-linked oligosaccharides. Consequently, Lecl cells are sensitive to the cytotoxic effects of the mannose-binding lectin Concanavalin A (Con A). Lecl cells were co-transformed with human DNA from A43 1 cells and eukaryotic expression plasmids containing the bacterial neo gene by calcium phosphate/DNA-mediated transformation. Co-transformants were selected for resistance to Con A and G-418. DNA from a primary co-transformant was purified and used to transform Lecl cells, resulting in secondary co-transformants. Both primary and secondary co-transformants exhibited in vitro GlcNAc-TI-specific enzyme activity, DNA gel blot analysis indicated that secondary co-transformants contained both human and neo sequences.

Who reads Co-transformation of Lec1 CHO cells with N-acetylgucosaminyltransferase 1 activity and a selectable marker?

Author

James Ripka; Michael Pierce; Nevis Fregien

Publisher

John Wiley and Sons; Wiley (John Wiley & Sons); John Wiley & Sons Inc.; Wiley (ISSN 0730-2312)

Published

1990

Language

EN

Field

Biochemistry, Genetics and Molecular Biology (Life Sciences)