Abstract
To investigate the expression level of testis-specific calcium-binding protein CBP86-IV in normal and asthenozoospermic human sperm. The total RNA was extracted from human sperm, and target cDNA was obtained by reverse transcription-polymerase chain reaction. Then the cDNA was used for quantitative PCR analysis and cloned into the prokaryotic expression vector pET-28a, respectively. The fusion protein was induced and expressed as inclusion body which was used to produce the polyclonal antibody against TSCBP86-IV. The protein expression level of TSCBP86-IV from normal human sperm and idiopathic asthenozoospermic samples was detected by the purified antibody. The experimental results showed that the protein expression of TSCBP86-IV was reduced in idiopathic asthenozoospermia and consistent with the transcriptional changing tendency which was detected by quantitative PCR analysis. The stable and reliable change of TSCBP86-IV may be taken as a new molecular marker for clinical diagnosis of idiopathic asthenozoospermia.
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