Goat β-lactoglobulin Gene Cloning and High Expression in the Mammary Gland of Transgenic Mice
山羊β乳球蛋白基因的克隆及其在转基因小鼠乳汁中的高效表达 Goat β-lactoglobulin Gene Cloning and High Expression in the Mammary Gland of Transgenic Mice

To clone goat beta-lactoglobulin (BLG) gene,two fragments were amplified from goat genomic DNA by LD-PCR method. The fragments were inserted in T-vectors before being spliced into the whole 7.2 kb BLG gene at a single restriction enzyme site of NarI. Consequently, the eukaryotic expression vector was constructed. All the clones were proved to be correct by restriction enzyme cutting and sequencing analysis. Six Founders (3 male symbol, 3 female symbol) of goat BLG transgenic mice were obtained by microinjection and BLG genes integration were confirmed by both PCR and Southern blot analyses. The milk was collected from two lactating female transgenic mice and goat BLG protein contents were measured with ELISA. The results showed that goat BLG protein in milk of the two mice were 23.49 mg/mL and 2.19 mg/mL, respectively.