[Expression and subcellular localization of the ORF4 gene of barley yellow dwarf virus GAV strain in baculovirus-insect cell expression system].
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.
Xia
Zong-Liang
ZL
College of Life Sciences, Henasn Agricultural University, Zhengzhou 450002, China. zongliangxia@yahoo.com
Wang
Mei-Ping
MP
Liu
Quan-Jun
QJ
Wang
Dao-Wen
DW
D000595:Amino Acid Sequence; D000818:Animals; D016367:Baculoviridae / Q000235:genetics / Q000378:metabolism*; D017343:Genes, Plant / Q000235:genetics*; D049452:Green Fluorescent Proteins / Q000235:genetics; D007313:Insecta / Q000235:genetics / Q000378:metabolism*; D017793:Luteovirus / Q000235:genetics*; D008969:Molecular Sequence Data; D016366:Open Reading Frames / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism
D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins