| dc.creator |
Niblett, C. L. |
|
| dc.creator |
Lee, R. F. |
|
| dc.creator |
Cevik, B. |
|
| dc.date |
2008-02-01T01:00:00Z |
|
| dc.date.accessioned |
2021-12-03T11:53:47Z |
|
| dc.date.available |
2021-12-03T11:53:47Z |
|
| dc.identifier |
b20e77f2-cfce-40b7-acf8-b380c20e6b80 |
|
| dc.identifier |
10.1007/s00705-007-1060-8 |
|
| dc.identifier |
https://avesis.sdu.edu.tr/publication/details/b20e77f2-cfce-40b7-acf8-b380c20e6b80/oai |
|
| dc.identifier.uri |
http://acikerisim.sdu.edu.tr/xmlui/handle/123456789/94347 |
|
| dc.description |
Expression of the RNA-dependent RNA polymerase (RdRp) of Citrus tristeza virus (CTV) was studied in vivo and in vitro using a polyclonal antiserum raised against the recombinant CTV-RdRp protein. Although a 57-kDa CTV-RdRp was expected to be expressed by a +1 translational frameshift at the carboxyl terminus of a 400-kDa polyprotein, a 50-kDa protein was detected in CTV-infected but not in healthy citrus tissue by Western blot. This suggests that the RdRp was cleaved from the CTV polyprotein. The 50-kDa protein was present in both the cytoplasmic and membrane fractions, but it accumulated mainly in the membrane fraction, where most of the replication-associated proteins of RNA viruses are found. When the expression of a cloned CTV-RdRp gene encoding a 60-kDa fusion protein was studied in vitro in a rabbit reticulocyte lysate system, two smaller proteins of about 50-kDa and 10-kDa were detected in addition to the expected 60-kDa protein. All three proteins were immunoprecipitated with the anti-CTV-RdRp serum, suggesting that the 50-kDa and 10-kDa proteins were fragments of the 60-kDa CTV-RdRp fusion protein. When the expression of the RdRp was analyzed at different times during in vitro translation, the 60-kDa and 50-kDa proteins were detected at all time points, and a small amount of the 10-kDa protein was detected after 30 min of translation. These results suggest that the CTV-RdRp may also be cleaved in vitro in the rabbit reticulocyte lysate. |
|
| dc.language |
eng |
|
| dc.rights |
info:eu-repo/semantics/closedAccess |
|
| dc.title |
In vivo and in vitro expression analysis of the RNA-dependent RNA polymerase of Citrus tristeza virus |
|
| dc.type |
info:eu-repo/semantics/article |
|