[Goat beta-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.
Gao
Jian-Jun
JJ
Institute of Medical Genetics, Shanghai Jiaotong University, Shanghai 200040, China.
Yan
Jing-Bin
JB
Huang
Ying
Y
Zeng
Yi-Tao
YT