The relationship between outer membrane protein D_2 of Pseudomonas aeruginosa and imipenem resistance
- VernacularTitle:铜绿假单胞菌外膜蛋白D_2与亚胺培南耐药的关系
- Author:
Chun-Xia GUO
;
Yong-Wen HE
;
Yan-Feng PAN
;
Shu-Li LI
;
Hua WANG
;
- Publication Type:Journal Article
- Keywords:
Pseudomonas aeruginosa;
Outer membrane protein D_2;
Polyclonal antibody;
Imipenem;
Resistance
- From:
Chinese Journal of Infection and Chemotherapy
2007;0(05):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective To prepare specific polyclonal antibodies to outer membrane protein (Opr) D_2 of Pseudomonas aeruginosa (PA),and explore the relationship between loss of OprD_2 and imipenem resistance.Methods The genomic DNA of PA was ex- tracted with phenol:chloroform.OprD_2 coding gene was amplified by PCR and prokaryotic expression vector pRSET-OprD_2 was constructed.OprD_2 protein was expressed by IPTG induction in E.coli BL21(DE3),and purified with SDS-PAGE.The new protein band was recovered and used as antigens to subcutaneously immunize two New Zealand rabbits to prepare poly- clonal antibody.The specificity of the antibody was determined by Western blot.The expression of OprD_2 in 32 clinical isolates of PA was detected with the prepared polyclonal antibody by Western blot.Results The vector pRSET-OprD_2 has been success- fully expressed in E.coli BL21 (DE3).The polyclonal anti-OprD_2 antibody with high specificity has been successfully pre- pared.Present results show that of the 27 imipenem-resistant PA clinical isolates,OprD2 protein was low-expressed in 5 iso- lates (18.5%) and normally expressed in 2 isolates (7.4%) but not expressed in 20 isolates (74.1%).Conclusions The loss or low-expression of OprD_2 is one of the essential mechanisms accounting for imipenem resistance in clinical isolates of PA.
