关键词: TAFA2基因, 脑组织特异表达基因, 基因剔除

Abstract: Objective To establish TAFA2 gene knockout mouse model and to create the condition for further in vivo study of its biological function and its role of development and growth in central nervous system. Methods The mouse genomic DNA sequence of TAFA2 gene was verified through bioinformatic analysis. According to the TAFA2 genomic DNA sequence, the strategy of gene targeting and constructing of knockout vector (pBR322-MK-MCS-TAFA2) were established. The knockout region spans from exon 4 to 5 of TAFA2 gene in mouse genome. The gene knockout vector pBR322-MK-MCS-TAFA2 was constructed and confirmed by restriction enzyme digestion and sequencing.Electroporation of ES cells with pBR322-MK-MCS-TAFA2 and screening of both G418 and Ganciclovoir resistant clones were performed according to common protocol. The homologous recombined ES cell clones were identified by PCR. Results After transplantation of homologous recombined ES cells into blastocysts through microinjection and mating of chimeras with C57BL/6J mice, 41 offspring with Aguoti fur in color were acquired, 14 of them (34.1%) show heterozygous genotype for TAFA2. Some heterozygote mice were intercrossed to generate mutant homozygotes. Finally, the cohort of mutant homozygotes for TAFA2 were established. Any abnormal change was not found by preliminary observa-tion of phenotype for heterozygous and homozygous mice. Conclusions The TAFA2 gene knockout mice have been established. The embryonic lethality in homologous mutant mice was not occurred.

Key words: TAFA2 gene, Gene expressed in specific brain tissues, Gene knockout