Molecular cloning of a novel rat gene Tsarg1, a member of the DnaJ/HSP40 protein family.
Beginning with a mouse gene mTSARG3, which was related to apoptosis of spermatogenic cells, bioinformatics was applied and a predicted novel rat gene full-length cDNA sequence was attained. Gene-specific primers were designed for PCR in rat testis cDNA library. A new gene Tsarg1 (GenBank Accession No. AY380804) was cloned, which is related to apoptosis in rat spermatogenic cells. The gene whose full cDNA length is 1176 bp containing 8 exons and 7 introns is located in rat chromosome 1q32-1q33, which encoded a protein containing 316 amino acid residues and being a new member of HSP40 protein family since the sequence contains the highly conserved J domain, which is present in all DnaJ-like proteins and is supported to have a critical role in DnaJ-DnaK protein-protein interactions. The results of RT-PCR and Northern blot analysis showed that Tsarg1 was specifically expressed in rat testis, which probably inhibits rat testis spermatogenic cell apoptosis.
Yang
Hong-Mei
HM
Human Reproductive and Stem Cell Engineering Institute, Central South University, Changsha, P. R. China.
Liu
Gang
G
Nie
Zhao-Yan
ZY
Nie
Dong-Song
DS
Deng
Yun
Y
Lu
Guang-Xiu
GX
D000595:Amino Acid Sequence; D000818:Animals; D017209:Apoptosis; D051017:Apoptosis Regulatory Proteins; D001483:Base Sequence; D015152:Blotting, Northern; D019556:COS Cells; D002522:Chlorocebus aethiops; D003001:Cloning, Molecular; D019295:Computational Biology / Q000379:methods; D017931:DNA Primers / Q000737:chemistry; D018076:DNA, Complementary / Q000378:metabolism; D004926:Escherichia coli / Q000378:metabolism; D005091:Exons; D020224:Expressed Sequence Tags; D005260:Female; D005982:Glutathione Transferase / Q000378:metabolism; D049452:Green Fluorescent Proteins / Q000378:metabolism; D050956:HSP40 Heat-Shock Proteins / Q000378:metabolism*; D006360:Heat-Shock Proteins / Q000235:genetics* / Q000502:physiology*; D007438:Introns; D008297:Male; D051379:Mice; D018832:Molecular Chaperones / Q000235:genetics* / Q000502:physiology*; D008969:Molecular Sequence Data; D009845:Oligospermia; D016366:Open Reading Frames; D010957:Plasmids / Q000378:metabolism; D017434:Protein Structure, Tertiary; D051381:Rats; D011993:Recombinant Fusion Proteins / Q000737:chemistry; D011994:Recombinant Proteins / Q000737:chemistry; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017386:Sequence Homology, Amino Acid; D013091:Spermatogenesis; D013737:Testis / Q000378:metabolism; D014018:Tissue Distribution; D014162:Transfection
D051017:Apoptosis Regulatory Proteins; D017931:DNA Primers; D018076:DNA, Complementary; D005982:Glutathione Transferase; D049452:Green Fluorescent Proteins; D050956:HSP40 Heat-Shock Proteins; D006360:Heat-Shock Proteins; D018832:Molecular Chaperones; D011993:Recombinant Fusion Proteins; D011994:Recombinant Proteins; C477075:DNAJB13 protein, human; C511342:Dnajb13 protein, rat; C531170:DNAJB13 protein, mouse