Stable expression of maize auxin-binding protein in insect cell lines.
To achieve continuous expression of the major maize auxin-binding protein (ABP1) in insect cells, the ABP1 gene coding region was placed under control of a baculovirus immediate-early gene promoter and transfected into Spodoptera frugiperda Sf9 cells. The ABP1 gene was detected in twelve cell lines, one of which was selected for detailed analysis. Immunolocalisation demonstrated that ABP1 was targeted to and retained in the endoplasmic reticulum (ER), in accordance with its signal peptide and carboxy-terminal KDEL ER-retention signal. We discuss the advantages of stable-transformation over transient expression systems for characterising proteins targeted to the secretory system of insect cells.
Henderson
J
J
School of Biological and Molecular Sciences, Oxford Brookes University, UK.
Atkinson
A E
AE
Lazarus
C M
CM
Hawes
C R
CR
Napier
R M
RM
Macdonald
H
H
King
L A
LA
D000818:Animals; D016367:Baculoviridae / Q000235:genetics; D001483:Base Sequence; D002461:Cell Line, Transformed; D018506:Gene Expression Regulation, Plant; D005822:Genetic Vectors; D007210:Indoleacetic Acids; D008969:Molecular Sequence Data; D009838:Oligodeoxyribonucleotides; D010937:Plant Growth Regulators; D010940:Plant Proteins / Q000096:biosynthesis / Q000235:genetics*; D011956:Receptors, Cell Surface / Q000096:biosynthesis / Q000235:genetics*; D018411:Spodoptera; D014170:Transformation, Genetic; D003313:Zea mays / Q000235:genetics*
D007210:Indoleacetic Acids; D009838:Oligodeoxyribonucleotides; D010937:Plant Growth Regulators; D010940:Plant Proteins; D011956:Receptors, Cell Surface; C046131:auxin receptor, plant