关键词: 慢性髓性白血病(CML), bcr启动子, 增强绿色荧光蛋白(EGFP), 转基因小鼠

Abstract: Objective To establish the bcr promoter drived green fluorescerce protein(bcr-EGFP) transgenic mouse model and to analyze the specificity of bcr promoter for further in vivo study. Methods The 1.1 kb bcr promoter was obtained from the genome of K562 cells (chronic myeloid leukemia cell line), at the same time, two restriction sites of Ase I、EcoR I were introduced into the up and down streams of this promoter. The CMV promoter of pEGFP-N1 plasmid was replaced by 1.1 kb bcr promoter to restructure pbcr-EGFP plasmid. Transfect the new recombinant plasmid into K562 cells and NIH3T3 cells by liposomes, after 24 hours, the expression was observed with fluorescentce. The results showed that this vector was constructed successfully. The bcr-EGFP recombinant fragment was obtained from pbcr-EGFP plasmid, and used to establish bcr-EGFP transgenic mouse by microinjection and the integration of target gene was detected by PCR. Results Total of 583 eggs of C57BL/6 were microinjected with the bcr-EGFP fragment. These eggs were transferred into 30 foster female mice, and 26 of which were pregnant and 90 pups were obtained, among which 3 pups (1♀, 2♂) were transgenic mice. Conclusion The successful establishment of bcr-EGFP transgenic mouse model has paved the way for future study of the specificity of bcr promoter.

Key words: Chronic myelogenous leukemia(CML), bcr promoter, Green fluorescence protein(EGP), Transgenic mouse