[Expression, purification and antibody preparation of recombinat SARS-CoV X5 protein].
X5 protein is one of the putative unknown proteins of SARS-CoV. The recombinant protein has been successfully expressed in E. coli in the form of insoluble inclusion body. The inclusion body was dissolved in high concentration of urea. Affinity Chromatography was preformed to purify the denatured protein, and then the product was refolded in a series of gradient solutions of urea. The purified protein was obtained with the purity of > 95% and the yield of 93.3 mg x L(-1). Polyclonal antibody of this protein was obtained, and Western blotting assay indicated that the X5 protein has the strong property of antigen. Sixty-eight percent of the recombinant protein sequence was confirmed by LC-ESI-MS/MS analysis.
Wang
Li-Na
LN
Institute of Materia Medica, Chinese Academy of Medical Sciences and Peking Union Medical College, Key Laboratory of Biosynthesis of Natural Products, Ministry of Health of PRC, Beijing 100050, China.
Kong
Jian-Qiang
JQ
Zhu
Ping
P
Du
Guan-Hua
GH
Wang
Wei
W
Cheng
Ke-Di
KD
D000595:Amino Acid Sequence; D000818:Animals; D000906:Antibodies / Q000302:isolation & purification*; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D015967:Gene Expression Regulation, Viral; D002479:Inclusion Bodies / Q000737:chemistry / Q000378:metabolism; D008969:Molecular Sequence Data; D011817:Rabbits; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification; D045473:Severe acute respiratory syndrome-related coronavirus / Q000235:genetics*; D014764:Viral Proteins / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification
D000906:Antibodies; D011994:Recombinant Proteins; D014764:Viral Proteins