Application of a nuclear localization signal gene in transgene mice  

作　　者：Jingpu Zhang Yuge Wang Guangsan Li Yu Shen Shaoxia Zhu Miao Du 

机构地区：[1]Chinese Acad Sci, Inst Dev Biol, Beijing 100080, Peoples R China

出　　处：《Chinese Science Bulletin》2002年第3期207-210,共4页

基　　金：This work was supported by the Special Support Grant of the Chinese Academy of Sciences (Grant No. STZ-3-05).

摘　　要：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 generatingEfficient gene transfer by cytoplasm co-injection will offer a powerful means for transgenic animals. Using co-injection in cytoplasm, two independent gene constructs, including bovine α-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. Expression 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 bioreactors 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 transgenic animals, which may be able to substitute the routine method of pronucleus-injection of fertilized eggs.

关 键 词：NUCLEAR localization signal CYTOPLASM injection trans-gene MOUSE GRANULOCYTE colony-stimulating factor integration and expression. 

分 类 号：Q781[生物学—分子生物学]