Design and preparation of non-tagged Yersinia pestis LcrV antigen in Escherichia coli and its immunogenicity in BALB/c mice.
The whole encoding sequence for Yersinia pestis LcrV antigen was cloned into pET-32a(+) and expressed in Escherichia coli BL21 (DE3). The LcrV was high level expressed in the E. coli cytoplasm in a completely soluble form. Recombinant LcrV could be purified from the supernatant of the bacteria lysate after chromatography using a combination of Phenyl-Sepharose F F, DEAE-Sepharose F F and Hiload Superdex 75. The final yield of approximately 3 g of purified rLcrV from 42 L bioreactor containing 25 L LB medium was obtained. High-titer IgG directed against rLcrV was detected positive after immunization on the BALB/c mice. The results presented here exhibit the ability to generate multi-gram quantities of non-tagged rLcrV from E. coli that can be used for the development of vaccine for preventing plague.
li
Jianmin
J
State Key Laboratory of Pathogens and Biosecurity, Laboratory of Applied Molecular Biology, Beijing Institute of Microbiology and Epidemiology, 20 FengTai Dongdajie Street, Beijing 100071, PR China. ljmqz@yahoo.com.cn
Li
Bing
B
Li
Guanlin
G
Ren
Jun
J
Zhang
Jinlong
J
Xu
Chun'e
C
Yang
Xiuxu
X
Liu
Shuling
S
Fu
Ling
L
Chen
Wei
W
D000818:Animals; D000907:Antibodies, Bacterial / Q000276:immunology; D000942:Antigens, Bacterial / Q000096:biosynthesis* / Q000276:immunology* / Q000302:isolation & purification; D015153:Blotting, Western; D004797:Enzyme-Linked Immunosorbent Assay; D004926:Escherichia coli / Q000378:metabolism*; D005822:Genetic Vectors; D051379:Mice; D008807:Mice, Inbred BALB C; D052899:Pore Forming Cytotoxic Proteins / Q000096:biosynthesis* / Q000276:immunology* / Q000302:isolation & purification; D021122:Protein Subunits / Q000096:biosynthesis / Q000302:isolation & purification; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000276:immunology / Q000302:isolation & purification; D020539:Sequence Analysis, Protein; D015010:Yersinia pestis / Q000378:metabolism*
D000907:Antibodies, Bacterial; D000942:Antigens, Bacterial; C043908:LcrV protein, Yersinia; D052899:Pore Forming Cytotoxic Proteins; D021122:Protein Subunits; D011994:Recombinant Proteins