Objective To explore the regulatory effect of fragile X mental retardation protein （FMRP） on the translation of microtubule associated protein 1B （MAP1B）. Methods The expressions of MAP1B protein and MAP1B mRNA in the brains of 1-week and 6-week old fragile X mental retardation-1 （FmrI） knockout （KO） mice were investigated by immunohistochemistry, Western blot, and in situ hybridization, with the age-matched wild type mice （WT） as controls. Results The mean optical density （MOD） of MAP1B was significantly decreased in each brain region in KO6W compared with WT6W, whereas in KO1W, this decrease was only found in the hippocampus and cerebellum. MAP1B in 6-week mice was much less than that in 1-week mice of the same genotype. The results of Western blot and in situ hybridization showed that MAP1B protein and MAP1B mRNA were significantly decreased in the hippocampus of both KO1W and KO6W. Conclusion The decreased MAP1B protein and MAP1B mRNA in the Fmrl knockout mice indicate that FMRP may positively regulate the expression of MAP1B.
目的:应用携带有报告基因-βGal的质粒对于使用L ipofectam ine 2000转染大鼠肝脏细胞及中国仓鼠肾脏成纤维细胞条件进行优化。方法:在96孔板上按不同比例混合L ipofectam ine 2000和质粒DNA后转染大鼠肝脏细胞及中国仓鼠肾脏成纤维细胞,成功转染的细胞因可表达β-Gal可分解底物X-gal在染液的存在下可使细胞呈蓝色,故计算蓝色细胞的比例即可了解转染的效率。应用方差分析的方法进行统计分析。结果:对于BHK细胞,当DNA与L ipofectam ine 2000的比例为0.27∶0.8(μg∶μL)时转染效率最高,为(43.2±9.3)%;对于BRL细胞,DNA与L ipofectam ine 2000的比例为0.10∶0.3(μg∶μL)时转染效率最高,可达(5.7±1.3)%。结论:L ipofectam ine 2000可有效转染哺乳动物细胞,不同细胞有不同的最佳转染条件。
