Expression, purification and characterization of the Lily symptomless virus coat protein from Lanzhou Isolate.
BACKGROUND
Lily symptomless virus (LSV) is widespread in many countries where lily are grown or planted, and causes severe economic losses in terms of quantity and quality of flower and bulb production. To study the structure-function relationship of coat protein (CP) of LSV, to investigate antigenic relationships between coat protein subunits or intact virons, and to prepare specific antibodies against LSV, substantial amounts of CP protein are needed.


RESULTS
Thus, full-length cDNA of LSV coat protein was synthesized and amplified by RT-PCR from RNA isolated from LSV Lanzhou isolate. The extended 33.6 kDa CP was cloned and expressed prokaryoticly and then purified by Ni-ion affinity chromatography. Its identity and antigenicity of recombinant CP were identified on Western-blotting by using the prepared anti-LSV antibodies.


CONCLUSIONS
The results indicate that fusion CP maintains its native antigenicity and specificity, providing a good source of antigen in preparation of LSV related antibodies. Detailed structural analysis of a pure recombinant CP should allow a better understanding of its role in cell attachment and LSV tropism. This investigation to LSV should provide some specific antibodies and aid to development a detection system for LSV diagnostics and epidemiologic surveys.
Cold and Arid Regions Environmental and Engineering Research Institute, Chinese Academy of Sciences, Lanzhou 730000, China.
Ruoyu
R
Wang
Guangpeng
G
Wang
Qi
Q
Zhao
Yu
Y
Zhang
Lizhe
L
An
Yun
Y
Wang
D000914:Antibodies, Viral / Q000276:immunology; D000956:Antigens, Viral / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification*; D015153:Blotting, Western; D036022:Capsid Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification*; D017788:Carlavirus / Q000235:genetics*; D002846:Chromatography, Affinity; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics / Q000302:isolation & purification; D015870:Gene Expression; D012367:RNA, Viral / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification; D020133:Reverse Transcriptase Polymerase Chain Reaction
D000914:Antibodies, Viral; D000956:Antigens, Viral; D036022:Capsid Proteins; D018076:DNA, Complementary; D012367:RNA, Viral; D011993:Recombinant Fusion Proteins