[Construction and activities  of suilysin mutants].
OBJECTIVE
To construct the suilysin mutant without hemolytic activity and evaluate its functions.


METHODS
The proline in 353 site of suilysin was site-directed mutated to alanine,  leucine and valine,  respectively. The recombinant mutants were renaturated  and purified by immobilized metal ion affinity chromatography,  and  the   purified proteins were evaluated in  the hemolytic activity and immunogenicity.


RESULTS
We obtained three mutants,  SLY(P353A),    SLY(P353L) and SLY(P353V). The SLY(P353V) mutant had   non-hemolytic activity. Western blotting and animal experiments showed that SLY(P353V) mutant still had immunogenicity.


CONCLUSIONS
Suilysin mutant SLY(P353V) has no hemolytic activity but remains immunogenicity.
College of Life Science, Sichuan Normal University, Chengdu, China. renzhiq@qq.com
Zhi-qiang
ZQ
Ren
Yu-ling
YL
Zheng
Shu-zhen
SZ
Gan
Qing-yu
QY
Lv
Huai-jie
HJ
Hao
Yong-qiang
YQ
Jiang
Hao
H
Zong
D000818:Animals; D001483:Base Sequence; D006460:Hemolysin Proteins / Q000235:genetics* / Q000276:immunology* / Q000302:isolation & purification; D006461:Hemolysis; D051379:Mice; D016297:Mutagenesis, Site-Directed; D009154:Mutation; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification; D013290:Streptococcal Infections / Q000401:mortality / Q000517:prevention & control; D017011:Streptococcus suis / Q000235:genetics / Q000276:immunology
D006460:Hemolysin Proteins; D011994:Recombinant Proteins; C104760:suilysin