Expression and subcellular localization of the ORF4 gene of barley yellow dwarf virus GAV strain in baculovirus-insect cell expression system.
- Author:
Zong-Liang XIA
1
;
Mei-Ping WANG
;
Quan-Jun LIU
;
Dao-Wen WANG
Author Information
1. College of Life Sciences, Henasn Agricultural University, Zhengzhou 450002, China. zongliangxia@yahoo.com
- Publication Type:Journal Article
- MeSH:
Amino Acid Sequence;
Animals;
Baculoviridae;
genetics;
metabolism;
Genes, Plant;
genetics;
Green Fluorescent Proteins;
genetics;
Insecta;
genetics;
metabolism;
Luteovirus;
genetics;
Molecular Sequence Data;
Open Reading Frames;
genetics;
Recombinant Fusion Proteins;
biosynthesis;
genetics;
metabolism
- From:Chinese Journal of Biotechnology
2007;23(6):1107-1111
- CountryChina
- Language:Chinese
-
Abstract:
According to published nucleotide sequences, ORF4 gene of barley yellow dwarf virus GAV (BYDV-GAV) was synthesized by reverse transcription-polymerase chain reaction (RT-PCR). The BYDV-GAV ORF4 gene was expressed in baculovirus -insect cell expression system efficiently, and western bolt analysis confirmed its expression product. Confocal laser scanning microscopy showed that GFP: ORF4 fusion protein was associated with the nuclear envelope of insect cells. By expressing the N- and C-terminal regions of ORF4-encoding product (P4) in insect cells combined with structure prediction, it was found that the N-terminal region of P4 containing four a-helices is required for targeting P4 to the nuclear envelope. These results provide a base for biological function of ORF4 gene during systemic infection of BYDV-GAV in host plants further.
