2.7.7.43 - 706038 2.7.7.43 a series of deletions from the 3'-end of the CMP-NeuAc synthetase coding region is constructed and expressed in Escherichia coli 662202 2.7.7.43 cloned in Escherichia coli as a His6-tagged fusion protein 672738 2.7.7.43 cloned in Escherichia coli XL-10 671567 2.7.7.43 cloning is achieved by complementation of the Chineses hamster ovary lec32 mutation that causes a deficiency in CMP-N-acetylneuraminate synthetase activity, it also causes polysialic acid to be expressed in the capsule of the CMP-Neu5Ac synthetase negative Escherichia coli mutant EV5 645273 2.7.7.43 DmCSAS is cloned into the baculovirus vector pBlueBac4.5 (pBlueBac-DmCSAS4) which is used to transfect LEC29.Lec32 cells 674570 2.7.7.43 DNA and amino acid sequence determination and analysis, when expressed in Drosophila S2 cells, DmCSS is mainly secreted into the culture medium, although partially detected in Golgi. Transient functional recombinant expression of myc-tagged wild-type and mutant enzymes in LEC29. Lec32 cells which lack a functional CSS, recombinant expression of His-tagged DmCSS in Escherichia coli 761215 2.7.7.43 DNA and amino acid sequence determination and analysis, when expressed in mammalian and insect cells, recombinant AaCSS shows in vivo and in vitro CSS activities. In contrast, when expressed in bacteria, it lacks CSS activity because the N-terminal hydrophobic region appears to induce protein aggregation, when expressed as C-terminally V5-tagged enzyme in Drosophila S2 cells, AaCSS is predominantly localized in the endoplasmic reticulum, but not in the Golgi. Transient functional recombinant expression of wild-type and mutant myc-tagged enzymes in LEC29. Lec32 cells which lack a functional CSS, recombinant expression of His-tagged AaCSS in Escherichia coli 761215 2.7.7.43 DNA and amino acid sequence determination and analysis, when expressed in mammalian and insect cells, recombinant TcCSS shows in vivo and in vitro CSS activities. In contrast, when expressed in bacteria, it lacks CSS activity because the N-terminal hydrophobic region appears to induce protein aggregation, when expressed in Drosophila S2 cells, TcCSS is predominantly localized in the endoplasmic reticulum, but not in the Golgi. Transient recombinant expression of myc-tagged enzyme in LEC29.Lec32 cells which lack a functional CSS, recombinant expression of His-tagged TcCSS in Escherichia coli 761215 2.7.7.43 expressed in CHO cells, HeLa cells, and NIH-3T3 cells 671794