Author/Authors :
Barzegar Yarmohammadi, Leila Avicenna Research Institute - Monoclonal Antibody Research Center, تهران, ايران , Barzegar Yarmohammadi, Leila tehran university of medical sciences tums - School of Medicine - Department of Medical Genetics, تهران, ايران , Modarresi, Mohammad Hossein tehran university of medical sciences tums - School of Medicine - Department of Medical Genetics, تهران, ايران , Modarresi, Mohammad Hossein Pasteur Iinstitute of Iran, تهران, ايران , Talebi, Saeed Avicenna Research Institute - Monoclonal Antibody Research Center, تهران, ايران , Talebi, Saeed tehran university of medical sciences tums - School of Medicine - Department of Medical Genetics, تهران, ايران , Hadavi, Reza Avicenna Research Institute - Monoclonal Antibody Research Center, تهران, ايران , Ostad Karampour, Mahyar Avicenna Research Institute - Monoclonal Antibody Research Center, تهران, ايران , Mahmoudi, Ahmad Reza Avicenna Research Institute - Monoclonal Antibody Research Center, تهران, ايران , Akhondi, Mohammad Mehdi Avicenna Research Institute - Reproductive Biotechnology Research Center, تهران, ايران , Rabbani, Hodjattallah Karolinska University Hospital - Karolinska Cancer Center - Immune and Gene Therapy Laboratory, Sweden. , Rabbani, Hodjattallah Avicenna Research Institute - Monoclonal Antibody Research Center, تهران, ايران , Jeddi-Tehrani, Mahmood Avicenna Research Institute - Monoclonal Antibody Research Center, تهران, ايران , Jeddi-Tehrani, Mahmood Karolinska University Hospital - Karolinska Cancer Center, - Immune and Gene Therapy Laboratory, Sweden
Abstract :
Introduction: Production of antibodies against specific proteins of testis germ cells isof great significance for the investigation of processes involved in spermatogenesis,study of infertility problems and determination of the probable role of these proteinsas cancer-testis antigens. Murine Testis Specific Recombinant Protein 101(mTEX101) is a 38kDa, GPI-anchored protein which is expressed in testis germ cellsof adult mice but it seems to be absent in other tissues. The structure and function ofmTEX101 is not completely understood yet, but it is speculated that it may transducebiochemical signals into the cytoplasm since mTEX101 does not have an intracellulardomain but the precise mechanisms are still ambiguous.Materials and Methods: RNA was extracted from three adult mice testis. The RNAwas used in RT-PCR, employing a pair of specific primers for mTEX101 ORFregion. TA-cloning technique was performed by the insertion of mTEX101 into apGEM-T Easy Vector, followed by its subcloning into a His-tagged expressionvector, pET-28a (+). The recombinant mTEX101 was then produced by transfectionof the expression vector into BL 21 (DE3) E. coli strain.Results: A recombinant protein, weighing 27kDa, was produced upon IPTGinductionof the bacterial host. The presence of mTEX101 protein was detectedthrough Western blot analysis by anti-mTEX101 peptide antibodies.Conclusion: We produced mTEX101 recombinant protein that could be used for theproduction of mono and polyclonal antibodies.
Keywords :
Antibody production , Gametogenesis , Germ cell , Glycoprotein , Recombinantprotein , Spermatogenesis , TES101 protein mouse , Testis.
