Study on induced expression and localization of LsGRP1 in the leaves of Lilium cv. Star Grazer
Date Issued
2007
Date
2007
Author(s)
Yao, Hsian-Wen
Abstract
In order to study expression of LsGRP1 protein in Lilum ‘Star Gazer’ leaves, LsGRP1 cDNA was constructed in protein expression vector of Escherichia coli. LsGRP1 expression and purification were carried on for preparation of LsGRP1 polyclonal antibody. Western blot analysis detected LsGRP1 as a protein of 38.2 kDa following extraction with non-ionic detergent Triton X-100 and reductant β-mercaptoethanol. A protein of 66 kDa was detected following extraction with hot water-NaOH and polyvinylpolypyrrolidone. Extracts of Lilum ‘Star Gazer’ leaves in different treatments were stably detected the protein of 38.2 kDa, but the protein of 66 kDa was only detected in the protein samples of systemic leaves, indicating that posttranslational modification(s) or large mixture composition of LsGRP1 might occur. In addition, the protein of 66 kDa could be detected in SA-treated protoplasts, and protein signal changes from the cell to culture supernatant as the time increased. Confocol microscopy study indicated that LsGRP1-GFP fusion protein was not present evenly in the protoplast cytoplasm, but accumulated inside the cell membrane. Thus, LsGRP1 may exist in the extracellular matrix or cell wall in Lilum ‘Star Gazer’ leaves was presumed.
Subjects
Lilum ‘Star Gazer’
polyclonal antibody
transient expression
protoplast
GFP fusion protein
confocol microscopy
File(s)![Thumbnail Image]()
Loading...
Name
ntu-96-R94633007-1.pdf
Size
23.53 KB
Format
Adobe PDF
Checksum
(MD5):0b49e5906b3c64f15de34d88ab0833bc