Construction of baculovirus transfer vectors for insect cell expression of human choline and ethanolamine kinase isoforms
Phosphatidylcholine and phosphatidylethanolamine are the most abundant phospholipid in eukaryotic cell membrane. Hwnan choline kinase and ethanolamine kinase play a very important role in the biosynthesis of phosphatidylcholine and phosphatidylethanolamine by catalyzing in the first phosphorylati...
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| Main Author: | |
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| Format: | Monograph |
| Language: | English |
| Published: |
Universiti Sains Malaysia
2008
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| Subjects: | |
| Online Access: | http://eprints.usm.my/48914/1/CHANG%20CHIA%20THAN%20-%2024%20pages.pdf http://eprints.usm.my/48914/ |
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| Summary: | Phosphatidylcholine and phosphatidylethanolamine are the most abundant phospholipid
in eukaryotic cell membrane. Hwnan choline kinase and ethanolamine kinase play a
very important role in the biosynthesis of phosphatidylcholine and
phosphatidylethanolamine by catalyzing in the first phosphorylation step of the pathway.
In this study, genes encoding hwnan choline kinase-P (hCK-P) and ethanolamine
kinase-2P (hEK-2P) isoform gene fragments were inserted into newly constructed
plasmid. The plasmid was constructed from commercially purchased transfer plasmid
pBAC4x-1. Open reading frames (ORF) ofhCK-P and hEK-2P were introduced into the
downstream of polyhedrin promoter for expression as histidine-tagged (his-tag) proteins.
The constructs were ready for homologous recombination with Autographa californica
Nuclear Polyhedrosis Virus (AcNPV) genome. Insect cells infected with the
recombinant AcNPV were expected to overexpress the hCK-P and hEK-2P proteins. |
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