Construction of Prokaryotic Expression Vectors of Antisense Nucleic Acid of LasR Gene and Its Effect on The Virulence of Pseudomonas aeruginosus
- VernacularTitle:铜绿假单胞菌LasR基因反义核酸原核表达载体的构建及对该菌毒力的影响
- Author:
Ling ZHANG
;
Junli ZHOU
;
Jingming LI
;
Fang LIAO
- Publication Type:Journal Article
- Keywords:
LasR gene, antisensenucleic acids, bacterial transformation, virulence factor
- From:
Progress in Biochemistry and Biophysics
2006;0(10):-
- CountryChina
- Language:Chinese
-
Abstract:
The LasR gene was amplified from the genome of Pseudomonas aeruginosus by PCR and recombined with plasmid pUCP18 reversely. The recombinant pUCP18/lasRantisense was verified with restriction analysis, PCR and sequence and was transformed in Pseudomonas aeruginosus. The biological effect of pUCP18/lasRantisense was detected by RT-PCR, NAD method and the assay of pyocyanin. The air tubes of rats were infected by pUCP18/lasRantisense strain and then carried on histopathologic slide check. Expected full length LasR fragment (721bp) can be extended from Pseudomonas aeruginosus gene with PCR technology. And it is consistent with LasR gene of Pseudomonas aeruginosa covered in GenBank (NO. NC_002516). The recombinant plasmid was constructed and transformed into Pseudomonas aeruginosus sucessfully. Compared with the rats which were infected by standard strain, the bronchitis of the rats which were infected by pUCP18/lasRantisense strain was obviously eased. It can be concluded that the antisensenucleic acid of LasR gene can depress the virulence of Pseudomonas aeruginosus and reveal a new target site for treatment.
