Construction and expression of RNase-resisting virus-like particles containing PSA mRNA
- VernacularTitle:内含人前列腺特异性抗原 mRNA 的耐核糖核酸酶病毒样颗粒的构建和表达
- Author:
Lu-Nan WANG
;
Jian-Ming WU
;
Jian-Ming PENG
;
Jin-Ming LI
;
Zhong-Fang WANG
;
Wei DENG
- Publication Type:Journal Article
- Keywords:
Prostate-specific antigen;
Virion;
Bacteriophages
- From:
Chinese Journal of Geriatrics
2001;0(03):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective To construct an expression system to produce the virus-like particles containing a part of the sequence of PSA mRNA, which are ribonuclease-resistant due to the encapsulation of the mRNA by bacteriophage MS2 coat proteins. Methods The PCR products of PSA cDNA fragments were cloned to TA vector pBS-T, then the targeted segments could be obtained when the pBS-T-PSA were digested by restriction endonuclease Hind Ⅲ and cloned to prokaryocytic expression vector pNCCL1. The recombinant plasmids named PNCCL1-PSA were transfected into E. Coli BL21-DE3 and induced to express with IPTG. Results The recombinant plasmids were successfully constructed. The bacteriophage MS2 coat protein which expressed in BL21 can self- assemble to form ribonuclease resistant virus-like particles and the PSA mRNA was encapsulated into virus-like particles. Conclusions The virus-like particle containing PSA mRNA can be expressed in prokaryocyte and it can be used as standard and control in detecting PSA mRNA. It provides a new, stable and ribonuclease-resistant RNA standard in RNA detection.
