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[Molecular cloning for testis spermatogenesis cell apoptosis related gene TSARG1 and Mtsarg1 and expression analysis
Jun-Jiang Fu1, Guang-Xiu Lu, Lu-Yun Li
1Laboratory of Human Reproductive Engineering, Central South University Xiangya Medical College, Changsha 410078, China.
Abstract:
Spermatogenesis cell apoptosis is a very complex process, which needs many molecules to take part in the programmable death of cells in testis. At present, research of apoptosis for spermatogenesis cell is at the primary step. It is very important to clone spermatogenesis cell apoptosis related genes and spermatogenesis genes in testis. Applying the bioinformatics and experiment technique, we have cloned human and mouse novel gene cDNA sequences--human testis and spermatogenesis cell apoptosis related gene 1 (TSARG1) and mouse testis and spermatogenesis cell apoptosis related gene 1(Mtsarg1) from human and mouse testis cDNA library respectively, using a cDNA fragment (GenBank accession number: BE644538) as an electronic probe, which was significantly changed in expression in cryptorchidism. The GeneBank accession numbers of Mtsarg1 and TSARG1 are AF399971 and AY032925 (NM_139073), respectively. The Mtsarg1 has a 55% identity and 61% similarity with TSARG1 at the amino acid level, which did not share significant homology with any other known protein in databases. The full-length cDNA of TSARG1 gene is 973 bp, including 549 bp open reading frame(ORF) and coding 183 amino acids, whereas the full-length cDNA of Mtsarg1 gene is 1103 bp, including 576 bp ORF and coding 192 amino acids. The predicted molecule weight of TSARG1 is 19948.61 Dolton, and the deduced iso-electric point is 10.24, whereas the Mtsarg1 is 20875.93 and is 9.83, being alkaline proteins. RT-PCR analysis showed that Mtsarg1 was expressed significantly in testis and faintly in epididymis in the ten tissues of testis, ovary, spleen, kidney, lung, heart, brain, epididymis, liver and skeletal muscle in mouse, while it wasn't expressed in the other eight tissues. Therefore, our results suggested that Mtsarg1 and TSARG1 would be pay potential roles in spermatogenesis cell apoptosis or spermatogenesis.
Insights
Researchers identified two novel genes, human testis and spermatogenesis cell apoptosis related gene 1 (TSARG1) and mouse Mtsarg1, crucial for male reproductive cell death and development. These genes show potential roles in spermatogenesis and apoptosis.
Area of Science:
- Reproductive Biology
- Molecular Genetics
- Cell Biology
Context:
- Spermatogenesis involves programmed cell death (apoptosis) regulated by numerous molecules.
- Research into spermatogenesis cell apoptosis is in its early stages.
- Identifying novel genes is critical for understanding male reproductive processes.
Purpose:
- To clone and characterize novel genes involved in spermatogenesis and apoptosis in human and mouse testis.
- To investigate the expression patterns of these newly identified genes.
Summary:
- Novel human (TSARG1) and mouse (Mtsarg1) testis and spermatogenesis cell apoptosis related genes were cloned using bioinformatics and experimental techniques.
- These genes exhibit significant homology to each other but not to known proteins, suggesting unique functions.
- Mtsarg1 is predominantly expressed in mouse testis and epididymis, indicating a role in male reproductive tissues.
Impact:
- The identification of TSARG1 and Mtsarg1 provides new molecular targets for studying spermatogenesis and male infertility.
- These findings contribute to a deeper understanding of the complex mechanisms underlying programmed cell death in male germ cells.
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