Abstract
A vector expressing human lysozyme (pBC1-hLYZ-GFP-Neo) was evaluated for gene and protein expression following liposome-mediated transformation of C-127 mouse mammary cancer cells. Cultures of G418-resistant clones were harvested 24-72 h after induction with prolactin, insulin and hydrocortisone. Target gene expression was analyzed by RT-PCR and Western blot and recombinant human lysozyme (rhLYZ) bacteriostatic activity was also evaluated. The hLYZ gene was correctly transcribed and translated in C-127 cells and hLYZ inhibited gram-positive bacterial growth, indicating the potential of this expression vector for development of a mammary gland bioreactor in goats. Guanzhong dairy goat skin fibroblasts transfected with pBC1-hLYZ-GFP-Neo were used to construct a goat embryo transgenically expressing rhLYZ by somatic nuclear transplantation with a blastocyst rate of 9.0 ± 2.8 %. These data establish the basis for cultivation of mastitis-resistant hLYZ transgenic goats.
| Original language | English |
|---|---|
| Journal | Biotechnology Letters |
| Volume | 34 |
| Issue number | 8 |
| Pages (from-to) | 1445-1452 |
| Number of pages | 8 |
| ISSN | 0141-5492 |
| DOIs | |
| Publication status | Published - Aug 2012 |
| Externally published | Yes |
Keywords
- Animals
- Animals, Genetically Modified
- Bioreactors
- Cell Line, Tumor
- Embryo, Mammalian
- Female
- Genetic Vectors
- Goats
- Green Fluorescent Proteins
- Humans
- Male
- Mammary Glands, Animal
- Mice
- Muramidase
- Polymerase Chain Reaction
- Recombinant Proteins
