Cloning and Analyzing Biological Activity of A2 Gene in Q? Phage
- VernacularTitle:Q?噬菌体A_2基因的克隆与生物活性分析
- Author:
Shu LI
;
Jie WANG
;
Tong WANG
- Publication Type:Journal Article
- Keywords:
Q? phage;
A2 gene;
Bacteriolysis activity
- From:
Journal of Medical Research
2006;0(06):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective To construct A2 gene expression vector in Q? phage by gene recombination technology, and then analyze its physiological activities. Methods Amplified A2 gene in Q? genome by PCR, cloned it into pBAD-24 expression vector to construct pBAD A2 recombinant plasmid. The recombinant plasmid was identified by restrictive enzymes digestion and DNA sequencing, then to be transfected into host cell JM109. After induced by Arabinose, the expression level of A2 was detected by SDS-PAGE. The growth curve of E.coli was obtained by phototurbidometry to test the bacteriolysis activity of pBADA2 in various host cells. Results After certified by PCR screening, DNA sequencing and restrictive enzymes digestion, the expression vector of pBADA2 was successfully constructed. The gene expression level is high in JM109 and related with Arabinose concentration, which reach its peak when Arabinose is 0.2%. OD660 value demonstrates that pBADA2 has the function of bacteriolysis, which could dissolve JM109、HB101 and 594 in E.coli rapidly, but not BE110. Conclusion The highly expressed vector pBADA2 was successfully constructed. The protein expressed has the ideal function of bacteriolysis. All of these provide theoretical and practical bases for developing new anti-bacteria drugs.
