Cloning of Human Ro60 cDNA and Construction of Baculovirus Expression Vector
- VernacularTitle:Ro60 cDNA的克隆及其重组杆状病毒表达载体的构建
- Author:
Lishao GUO
;
Jianzhong ZHANG
- Publication Type:Journal Article
- Keywords:
Gene duplication;
Genetic vectors;
Ro60
- From:
Chinese Journal of Dermatology
1994;0(02):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective To construct a baculovirus expression vector for expression of human Ro60 cDNA. Methods Ro60 cDNA was cloned from Hela cell RNA with RT-PCR and was inserted into plasmid pFastBac HTc. Plasmid pFastBac HTc-Ro60 was then transformed into E. coli DH10Bac competent cells for transposition. The constructed recombinant was determined by both blue-white colony screening and PCR analysis. Results A 1.56kb Ro60 cDNA was obtained from Hela cell RNA and was successfully inserted to plasmid pFastBac HTc. DNA sequencing and restriction enzyme analysis revealed that both open reading frame and sequence of the cDNA were identical to Ro60 sequence previously published. Specific transposition and virus recombination were obtained in the vector Bacmid-Ro60. Conclusion Ro60 cDNA has been successfully cloned and constructed to baculovirus expression vector, which is useful for protein expression and further study of this autoantigen in the pathogenesis of lupus erythematosus.
