[Cloning of human lysosomal acid beta-glucosidase gene and its expression in COS7 cells].
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.
Zhang
Yanli
Y
Center of Animal Embryo Engineering and Technology, Nanjing Agricultural University, Nanjing 210095, China.
Xu
Dan
D
Wang
Ziyu
Z
Meng
Li
L
Wang
Feng
F
D000818:Animals; D019556:COS Cells; D002522:Chlorocebus aethiops; D003001:Cloning, Molecular; D005822:Genetic Vectors / Q000235:genetics; D005962:Glucosylceramidase / Q000235:genetics* / Q000378:metabolism*; D049452:Green Fluorescent Proteins / Q000235:genetics; D006801:Humans; D008247:Lysosomes / Q000201:enzymology*; D011995:Recombination, Genetic; D014162:Transfection
D049452:Green Fluorescent Proteins; D005962:Glucosylceramidase