Cloning of human lysosomal acid beta-glucosidase gene and its expression in COS7 cells.
- Author:
Yanli ZHANG
1
;
Dan XU
;
Ziyu WANG
;
Li MENG
;
Feng WANG
Author Information
1. Center of Animal Embryo Engineering and Technology, Nanjing Agricultural University, Nanjing 210095, China.
- Publication Type:Journal Article
- MeSH:
Animals;
COS Cells;
Cercopithecus aethiops;
Cloning, Molecular;
Genetic Vectors;
genetics;
Glucosylceramidase;
genetics;
metabolism;
Green Fluorescent Proteins;
genetics;
Humans;
Lysosomes;
enzymology;
Recombination, Genetic;
Transfection
- From:
Chinese Journal of Biotechnology
2009;25(2):263-267
- CountryChina
- Language:Chinese
-
Abstract:
In this study, we amplified human lysosomal acid beta-glucosidase (GlcCerase) gene by RT-PCR from human placenta, and analyzed the sequence of the PCR product cloned in pMD-19T. The gene identity was 99% comparable to that of the reported human GlcCerase cDNA sequence in GenBank. The GlcCerase gene digested with Xho I was subcloned into eukaryotic express vector pEGFP-C1 to generate recombinant expression vector pEGFP-GlcCerase. After identified the recombinant plasmid by restriction enzyme digestion, we transfected pEGFP-GlcCerase into COS7 cells by liposome. GlcCerase mRNA was expressed and the activity of GlcCerase was also detected in COS7 cells. This study would lay a foundation for the function of GlcCerase and its production by transgenic bioreactor.
