Efficient gene transfer by cytoplasm co-injec-tion will offer a powerful means for transgenic animals. Us-ing co-injection in cytoplasm, two independent gene con-structs, including bovine a-s1-casein-hG-CSF and a mamm...Efficient gene transfer by cytoplasm co-injec-tion will offer a powerful means for transgenic animals. Us-ing co-injection in cytoplasm, two independent gene con-structs, including bovine a-s1-casein-hG-CSF and a mammal expression vector expressing a nuclear localization signal (mNLS), were introduced into fertilized mouse eggs. The target gene construct was docked into host nucleus probably by the nuclear localization signal. Transgene mice have been obtained at 58% (29/50) of integration ratio. Ex-pression level of the positive transgene mice was detected by Western blotting. Maximal expression of human G-CSF was estimated about 540 mg/L of milk. The expression ratio was up to 75% (9/12). The results here have important practical implications for the generation of mammary gland bioreac-tors and other transgene studies. Co-injection of a target gene with an expression vector of a mammal nuclear localization signal by cytoplasm appears to be a useful, efficient and easy strategy for generating展开更多
基金This work was supported by the Special Support Grant of the Chinese Academy of Sciences (Grant No. STZ-3-05).
文摘Efficient gene transfer by cytoplasm co-injec-tion will offer a powerful means for transgenic animals. Us-ing co-injection in cytoplasm, two independent gene con-structs, including bovine a-s1-casein-hG-CSF and a mammal expression vector expressing a nuclear localization signal (mNLS), were introduced into fertilized mouse eggs. The target gene construct was docked into host nucleus probably by the nuclear localization signal. Transgene mice have been obtained at 58% (29/50) of integration ratio. Ex-pression level of the positive transgene mice was detected by Western blotting. Maximal expression of human G-CSF was estimated about 540 mg/L of milk. The expression ratio was up to 75% (9/12). The results here have important practical implications for the generation of mammary gland bioreac-tors and other transgene studies. Co-injection of a target gene with an expression vector of a mammal nuclear localization signal by cytoplasm appears to be a useful, efficient and easy strategy for generating
