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POSTER NO: 222 Znf230 is a Mouse Spermatogenesis Candidate Gene
Weimin Qiu, Sizhong Zhang, Hui Wu Spermatogenesis is a complex process of cell development and differentiation during which extensive changes in cell morphology and intracellular organization occur. This process requires the highly regulated expression of a network of genes located on both the autosomes and the sex chromosomes. Recently, zinc finger genes, which belong to one of the largest gene families in mammals, were proved to be involved in their spermatogenesis. Human ZNF230, which is a novel ring finger gene, is cloned by mRNA differential display with testicular tissues of fertile adults and patient of azoospermia. Taking its mRNA sequence as reference, we cloned the mouse homolog Znf230 (AF353167) by two methods. 1. In silico by assembling the highly homologous ESTs existing in GenBank; 2. by rapid amplification of cDNA ends (RACE) to obtain the full length cDNA. As a result, a 982bp testis transcript which is also a ring finger gene message was gained. This cDNA and its predicated amino acid sequence are 93% and 98% homologous to those of human ZNF230 respectively. Multiple tissue northern blot analysis reveals that there are three transcripts of the gene. The 4.4kb and 2.4kb transcripts are ubiquitously expressed and the 1 kb transcript is specifically expressed in testis. The results of RT-PCR experiments indicated that the gene highly expressed in mouse testis reaches the adult level of transcription at the 21 postnatal day. In situ hybridization revealed that the mRNA of Znf230 is expressed in spermatogonia and spermatocytes. These results suggest that mouse Znf230, as its human homologue, is a testis-specific ring finger gene and may function during the process of differentiation from the spermatogonia to round spermatids. |