[Preparation of polyclonal antibody and prokaryotic expression of   recombinant protein RelA from Mycobacterium tuberculosis].
OBJECTIVE
To prepare polyclonal antibodies against RelA protein of Mycobacterium tuberculosis.


METHODS
RelA gene segment was inserted into pET-32a(+) and the recombinant protein RelA was expressed in E.coli under IPTG induction.The protein was purified and identified by SDS-PAGE and Western blot.Polyclonal antibody to RelA was got by immunizing rabbits with the protein. Quality and quantity of the antibody was identified.


RESULTS
RelA gene segment was successfully inserted into pET-32a(+) and recombinant protein RelA was obtained.The polyclonal antibody to RelA had a good specificity, and the titer reached more than 1:6 400.


CONCLUSIONS
RelA recombinant protein and rabbit anti-RelA polyclonal antibody with high specificity were obtained, which provided good tools for further studying functional characterization of RelA.
Department of Laboratory Medicine, Weifang Medical University, Weifang, China. yizhengjun@126.com
Zheng-jun
ZJ
Yi
Yu-rong
YR
Fu
Fu-liang
FL
Xu
Kun-shan
KS
Gao
Meng
M
Li
Jian-hua
JH
Li
D000818:Animals; D000906:Antibodies / Q000276:immunology*; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D009169:Mycobacterium tuberculosis / Q000235:genetics* / Q000276:immunology*; D011817:Rabbits; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification; D051996:Transcription Factor RelA / Q000235:genetics* / Q000276:immunology* / Q000302:isolation & purification
D000906:Antibodies; D011994:Recombinant Proteins; D051996:Transcription Factor RelA