细胞凋亡关键蛋白FADD的组织分布分析

Tissue distribution analysis of key cell apoptosis protein FADD

目的:分析不同物种FADD的蛋白序列进化规律及FADD在不同组织中的转录和表达分布特征。方法:使用Mega 5.0软件对主要物种中FADD的蛋白序列进行比对分析,根据蛋白序列相似性使用邻近法绘制进化树。取FADD+/-小鼠脑、肝、肾、心、肺、肌肉、脾脏、胃、肠,分别提RNA并反转录成cDNA和同时制备相应的蛋白样品,将FADD+/-小鼠在核酸水平和蛋白水平各主要组织中FADD的表达结果与小鼠FADD表达的UniGene EST Profile结果进行综合比较分析。结果:获得不同物种FADD的蛋白序列比对结果及进化规律,确定FADD在小鼠肺、心脏、胃中相对高表达的可靠结果,其他组织中FADD的表达量还有待进一步验证。结论:FADD的蛋白序列在进化中相对比较保守,其在小鼠肺、心脏、胃组织中高表达,这为进一步研究FADD在不同组织中生物功能奠定了良好的基础。

Objective： To analyze the evolution of FADD protein in different species and its transcription and expression distribution characteristics in different tissues .Methods： Protein sequences of FADD in eight main species were compared and analyzed by using the software Mega 5.0 , and the neighbor-joining phylogenetic tree was drawn according to the protein sequence similarity .FADD＋/-mice were sacrificed and their brain , liver, kidney, heart, lung, muscle, spleen, stomach, intestine were obtained and then divided into two parts .Half of the tissues were used to extract RNA for RT-PCR, the rests were used for western blot analyses .The results of FADD expression in FADD＋/-mice were compared with the results of UniGene EST Profile at both mRNA and protein levels.Results： On the one hand, we obtained the protein sequence alignment and evolution law of FADD in different species.On the other hand, we found FADD expression was relatively high in lung , heart, stomach, while the expression of FADD in other tissues remains to be verified .Conclusion：The present studies find that FADD is highly conserved in the process of evolution and it is highly expressed in lung , heart and stomach .These results provide a foundation for further investigation of the biological function of FADD in different tissues .