M.A. Thesis:

cloning of sialyltransferase in factor IX producing S۲ cell line

Given the fact that it has been shown today that the insect expressive system has a broad ability to produce recombinant human proteins with proper folding and post-translational modifications, biologists are more likely to be interested in the past. However, the S2 expression system Expression systems are insects that have many applications, for example, can be similar to the addition of sugar units in the process of glycosylation of mammalian herbs, but it is incapable of adding galactose and sialic acid to secretion proteins. It is due to the ability of these cells And the importance of doing glycosylation Correct in recombinant human proteins, making changes in this system is essential for glycosylated recombinant protein synthesis, similar to that of mammals. In this study, the cloning of beta 1 and 4 galactosyl, alpha 2, and 6 of the transfer fluid of Pmt reagent and its transfection into S2-producing cells Recombinant IX factor was performed. After induction of these cells, the culture medium of these cells was collected at time intervals of 24, 48 and 72 hours, and the activity and expression of the secreted factor IX were studied using one-step coagulation and ELISA. The results showed that insertion of beta 1 and 4 galactosyl, alpha 2 and 6 sialyl transferase gene had no significant effect on the expression and activity of this coagulation protein.

Keywords: factor ix, hemophiliaT