Producing Recombinant mTEX101; a Murine Testis Specific Protein.
BACKGROUND
Production of antibodies against specific proteins of testis germ cells is of great significance for the investigation of processes involved in spermatogenesis, study of infertility problems and determination of the probable role of these proteins as cancer-testis antigens. Murine Testis Specific Recombinant Protein 101 (mTEX101) is a 38kDa, GPI-anchored protein which is expressed in testis germ cells of adult mice but it seems to be absent in other tissues. The structure and function of mTEX101 is not completely understood yet, but it is speculated that it may transduce biochemical signals into the cytoplasm since mTEX101 does not have an intracellular domain but the precise mechanisms are still ambiguous.


METHODS
RNA was extracted from three adult mice testis. The RNA was used in RT-PCR, employing a pair of specific primers for mTEX101 ORF region. TA-cloning technique was performed by the insertion of mTEX101 into a pGEM-T Easy Vector, followed by its subcloning into a His-tagged expression vector, pET-28a (+). The recombinant mTEX101 was then produced by transfection of the expression vector into BL 21 (DE3) E. coli strain.


RESULTS
A recombinant protein, weighing 27kDa, was produced upon IPTG-induction of the bacterial host. The presence of mTEX101 protein was detected through Western blot analysis by anti-mTEX101 peptide antibodies.


CONCLUSIONS
We produced mTEX101 recombinant protein that could be used for the production of mono and polyclonal antibodies.
Monoclonal Antibody Research Center, Avicenna Research Institute, ACECR, Tehran, Iran ; Department of Medical Genetics, School of Medicine, Tehran University of Medical Sciences, Tehran, Iran.
Barzegar Yarmohammadi
BY
Leila
Modarresi
M
Mohammad Hossein
Talebi
T
Saeed
Hadavi
H
Reza
Ostad Karampour
OK
Mahyar
Mahmoudi Ahmad
MA
Reza
Akhondi Mohammad
AM
Mehdi
Rabbani
R
Hodjattallah
Jeddi-Tehrani
JT
Mahmood
Antibody production; Gametogenesis; Germ cell; Glycoprotein; Recombinant protein; Spermatogenesis; TES101 protein mouse; Testis