Objectives: Previous data has reported similarity between human brain and testis gene expression patterns. Brain is the most important tissue in human speciation. Hence, it means that human testis could also play a crucial role in human speciation if these two tissues exhibit similar gene expression patterns. However, previous reports were based on only limited and scattered data. Determining the large scale anatomy of gene expression patterns of various human tissues could draw a more convincing conclusion, and better our understanding of the correlation/inter-correlation among different tissues. Furthermore, it could also provide a clue for evolutionary study. Methods: To obtain gene expression information for large-scale data analysis, expression data of 760 Unigenes in seventeen human tissues (liver, lung, testis, brain, ovary, uterus, colon, stomach, heart, eye, kidney, spleen, gall bladder, breast, thymus, prostate and pancreas) were retrieved by DDD (differential digital display) analysis, and this expression data was subjected to clustering analysis. These Unigenes represent a wide range of genes classified according to their characterization and function. Results: Among the 17 tissues, the highest similarity in gene expression patterns was between human brain and testis, based on DDD and clustering analysis. Genes contributing to the similarity include ribosomal protein (RP) genes as well as genes involved in transcription, translation and cell division. Conclusions: Present results provide evidence to support the proposal that human testis and brain share the highest similarity of gene expression patterns. The implications of the similarity regarding that both brain and testis contributed to human speciation are discussed.