Enhanced protein secretion from insect cells by co-expression of the chaperone calreticulin and translation initiation factor eIF4E

C.Y. Teng, S.L. Chang, M.M. van Oers, T.Y. Wu

Research output: Contribution to journalArticleAcademicpeer-review

7 Citations (Scopus)

Abstract

Host protein synthesis is shut down in the lytic baculovirus expression vector system (BEVS). This also affects host proteins involved in routing secretory proteins through the endoplasmic reticulum (ER)-Golgi system. It has been demonstrated that a secretory alkaline phosphatase–EGFP fusion protein (SEFP) can act as a traceable and sensitive secretory reporter protein in BEVS. In this study, a chaperone, calreticulin (CALR), and the translation initiation factor eIF4E were co-expressed with SEFP using a bicistronic baculovirus expression vector. We observed that the intracellular distribution of SEFP in cells co-expressing CALR was different from co-expressing eIF4E. The increased green fluorescence emitted by cells co-expressing CALR had a good correlation with the abundance of intracellular SEFP protein and an unconventional ER expansion. Cells co-expressing eIF4E, on the other hand, showed an increase in extracellular SEAP activity compared to the control. Utilization of these baculovirus expression constructs containing either eIF4E or CALR offers a significant advantage for producing secreted proteins for various biotechnological and therapeutic applications.
Original languageEnglish
Pages (from-to)68-78
JournalMolecular Biotechnology
Volume54
Issue number1
DOIs
Publication statusPublished - 2013

Keywords

  • baculovirus expression system
  • ribosome entry site
  • endoplasmic-reticulum
  • quality-control
  • messenger-rna
  • calnexin
  • infection
  • pathway
  • er
  • glycoprotein

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