[Prokaryotic expression of LMoV CP gene and preparation of its antiserum].
OBJECTIVE
To prepare antiserum against the CP of Lilg mottle virus (LMoV).


METHODS
Specific primer was designed according to Genbank to amplify CP gene of LMoV of Fritillaria thumbergii and its sequence was analyzed. Then the CP gene was inserted into pSBET and expressed in Escherichia coli BL21 (DE3) plys E strain. The objective protein was purified by 12% SDS-PAGE firstly and subsequently 5% - 20% gradient SDS-PAGE. The antiserum against the CP was raised in mouse and their specificity was determined by Western blot. The ability to combine with nature LMoV particles was confirmed by ELISA analysis.


RESULTS
LMoV CP gene shared 95% - 99% nucleotide identities and 98% - 100% amino acid identities with the CP genes reported on Genbank. The antiserum was special to LMoV CP and IgG against LMoV could combine LMoV particles.


CONCLUSIONS
The antiserum prepared in this study is suitable for LMoV detection.
Biological and Pharmaceutical Enginnering College, Western Anhui University, Lu'an 237012, China. weichuanbao@sina.com
Chuan-Bo
CB
Wei
Hou-Jun
HJ
Yao
Yu
Y
Yang
Chun-Yun
CY
Liu
D000595:Amino Acid Sequence; D000818:Animals; D000906:Antibodies / Q000276:immunology / Q000302:isolation & purification; D015153:Blotting, Western; D036022:Capsid Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology; D003001:Cloning, Molecular; D017931:DNA Primers; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D031410:Fritillaria / Q000235:genetics / Q000821:virology*; D005822:Genetic Vectors; D007106:Immune Sera / Q000276:immunology / Q000302:isolation & purification*; D051379:Mice; D017800:Potyvirus / Q000235:genetics* / Q000276:immunology; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology
D000906:Antibodies; D036022:Capsid Proteins; D017931:DNA Primers; D007106:Immune Sera; D011994:Recombinant Proteins