Description:
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hLZ goats were developed to produce human lysozyme (hLZ) in their milk, a protein which finds application in the food industry and in therapeutic applications.
These hLZ goats were generated through transfection of female primary goat fibroblasts followed by transfer into enucleated oocytes.
Two different constructs were used to generate independent goat lines expressing the human lysozyme gene (lyz) specifically in the mammary gland.
In the pBC-hLZ construct, the 4.8 kb lyz genomic DNA region was placed under the goat beta-casein promoter (P-csn2-CAPHI), followed by the 3’ regulatory sequence of beta-casein (T-csn2-CAPHI) and a neomycin resistance cassette driven by the CMV promoter.
The pBLG-hLZ construct was used to express lyz (4.8 kb genomic region) under the bovine beta-lactoglobulin promoter (P-lgb-BOVIN), followed by a bovine growth factor polyadenylation fragment (T-bgh-BOVIN). Here, a neomycin resistance cassette driven by the thymidine kinase promoter (P-thymidine kinase) was co-transfected with the lyz construct for transgene selection.
Transgenic lines were identified by PCR and Southern Blot analysis. Both lines of transgenic goats contain one copy of the transgene.
Sources:
Yu et al. (2013), Large-scale production of functional human lysozyme in transgenic cloned goats. Journal of Biotechnology 168:676-683.
Xu et al. (2021), An event-specific Real-Time PCR method for measuring transgenic lysozyme goat content in trace samples. Foods 10(5):925.
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Links regarding to this GMO:
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Scientific publication – Yu et al., (2013) https://www.sciencedirect.com/science/article/abs/pii/S0168165613004562?via%3Dihub,
Scientific publication – Xu et al., (2021) https://www.mdpi.com/2304-8158/10/5/925
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Transformation / Modification technique:
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Reproductive cloning
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