Construction and identification of recombinant lentiviral vector of hNoc4L gene.
- Author:
Tingting WANG
1
;
Shujuan WANG
;
Jinghua YAN
Author Information
1. CAS Key Laboratory of Pathogenic Microbiology and Immunology, Institute of Microbiology, Chinese Academy of Science, Beijing 100101, China.
- Publication Type:Journal Article
- MeSH:
Animals;
Cell Line;
Genetic Vectors;
genetics;
Lentivirus;
genetics;
metabolism;
Mice;
Nuclear Proteins;
biosynthesis;
genetics;
Promoter Regions, Genetic;
Recombinant Proteins;
biosynthesis;
genetics;
Saccharomyces cerevisiae Proteins;
biosynthesis;
genetics
- From:
Chinese Journal of Biotechnology
2010;26(11):1569-1575
- CountryChina
- Language:Chinese
-
Abstract:
Formation and nuclear export of pre-ribosomes requires many nucleolar complexes, hNoc4L which contains a conserved Noc doman is a homolog of nucleolar complex associated 4 (S. cerevisiae), but its function is completely unclear. Here, we successfully got the recombinant lentiviral vector p113.7-EF1-hNoc4L-Flag by replacing the U6 promoter in p113.7 with EF1alpha promoter, and then inserted hNoc4L to down-stream of the EF1alpha prompter. We determined the transduction efficiency in different mammalian cell lines based on lentiviral packaging system. Subsequently, we analyzed the immunogenicity of the recombinant lentivirus and stable expression of hNoc4L in RAW264.7 cells. The results showed that the recombinant lentivirus characterized a high transduction efficiency, long-term expression and low immunogenicity. Therefore, we pave the way for further identification of the biological activity of hNoc4L protein during ribosome biogenesis in mammalian.
