Producing Recombinant mTEX101; a Murine Testis Specific Protein

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.