[Recombinant expression, purification and adhesion function identify of Bacillus anthracis BslA(260 -652) protein].
OBJECTIVE
To obtain the recombinant BslA(260-652) protein of Bacillus anthracis and prepare its antibody for the adhesion activity studies.


METHODS
The fragment coding BslA(260-652) was cloned into pET28a(+) plasmid and induced to express recombinant protein in E. coli Rosetta (DE3) by Isopropyl beta-D-1-thiogalactopyranoside (IPTG). The expressed recombinant soluble protein was purified by a column packed with Ni Resin. Purified protein was used as the antigen to immunize BABL/c mice for three times to raise polyclonal antibody. The adhesion activity of BlsA(260-652) was detected by immunofluorescence experiments and bacterial adherence assays.


RESULTS
The purity of the purified soluble BslA(260-652) was about 87.4%. ELISA assay titer of antiserum from vaccinated mice reached 1:20000. Western blot showed the antiserum could specifically recognize endogenous BslA protein. The purified BslA(260-652) displayed a typical adhesion-like function. Either the anti-BslA serum or the BslA(260-652) protein could inhibit A16R's Hela adherence.


CONCLUSIONS
The recombinant BslA(260-652) protein was successfully obtained, which would lay the foundation for further research of the anthrax vaccine and the role of this S-layer protein in the pathogenesis of anthrax.
School of Life Sciences, Anhui University, Hefei 230039, China. makunwx@gmail.com
Kun
K
Ma
Yanchun
Y
Wang
Haoxia
H
Tao
Jie
J
Dong
Chen
C
Cao
Buchang
B
Zhang
Chunjie
C
Liu
D018829:Adhesins, Bacterial / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism*; D000818:Animals; D000881:Anthrax / Q000382:microbiology; D001408:Bacillus anthracis / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D002460:Cell Line; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D005260:Female; D015870:Gene Expression; D006801:Humans; D008562:Membrane Glycoproteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism*; D051379:Mice; D008807:Mice, Inbred BALB C; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism
D018829:Adhesins, Bacterial; D008562:Membrane Glycoproteins; D011994:Recombinant Proteins; C055764:S-layer proteins