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C‐terminal region of hTPO is important for secretion and expression in insect cells
Author(s) -
Man Hyea Kyung,
Chung Joo Young,
Park Sang Kyu,
Joo Semg Myung,
Park Seung Kook,
Koh Yeo Wook
Publication year - 1999
Publication title -
iubmb life
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.132
H-Index - 113
eISSN - 1521-6551
pISSN - 1521-6543
DOI - 10.1080/15216549900201803
Subject(s) - trichoplusia , secretion , biology , cell culture , baculoviridae , microbiology and biotechnology , amino acid , thrombopoietin , glycosylation , recombinant dna , spodoptera , biochemistry , gene , lepidoptera genitalia , genetics , botany , stem cell , haematopoiesis , noctuidae
Human thrombopoietin (hTPO) variant cDNAs truncated in the C‐terminal regions of wild‐type hTPO (332 amino acids) were constructed by PCR and expressed in Trichoplusia ni (Tn5) insect cells using a baculovirus expression system. Each variant, hTPO163 (amino acids 1‐163), hTPO198 (1‐198) and hTPO245 (1‐245), was produced in insect cells with very low efficiency in comparison with wild‐type hTPO. Immunoblot analysis showed that the predicted 20, 25 and 34 kDa molecular sizes corresponding to hTPO163, hTPO198 and hTPO245, respectively, were barely detected in culture medium and most of the proteins remained within the cell. These results suggest that C‐terminal regions containing potential N‐glycosylation sites of hTPO are required for the secretion of hTPO into culture medium as well as expression in insect cells.