[Molecular cloning for testis spermatogenesis cell apoptosis related gene TSARG1 and Mtsarg1 and expression analysis for Mtsarg1 gene].
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.
Fu
Jun-Jiang
JJ
Laboratory of Human Reproductive Engineering, Central South University Xiangya Medical College, Changsha 410078, China.
Lu
Guang-Xiu
GX
Li
Lu-Yun
LY
Liu
Gang
G
Xing
Xiao-Wei
XW
Liu
Shang-Feng
SF
D000595:Amino Acid Sequence; D000818:Animals; D017209:Apoptosis / Q000235:genetics*; D001483:Base Sequence; D002874:Chromosome Mapping; D002889:Chromosomes, Human, Pair 2 / Q000235:genetics; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000737:chemistry / Q000235:genetics; D015870:Gene Expression; D006801:Humans; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D008969:Molecular Sequence Data; D011506:Proteins / Q000235:genetics*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D012689:Sequence Homology, Nucleic Acid; D026801:Synteny; D013737:Testis / Q000166:cytology / Q000378:metabolism*
D018076:DNA, Complementary; D011506:Proteins; D012333:RNA, Messenger; C476406:SPATA3 protein, human