Shulin Shen1, Jinzi Wang, Jihong Liang, Chunhui Zhu. 1. Department of Andrology, The First Affiliated Hospital of Guangxi Medical University, Nanning, 530021, China, shenforest@163.com.
Abstract
OBJECTIVES: To investigate the expression level of testis-specific calcium-binding protein CBP86-IV in normal and asthenozoospermic human sperm. METHODS: 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. RESULTS: 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. CONCLUSIONS: The stable and reliable change of TSCBP86-IV may be taken as a new molecular marker for clinical diagnosis of idiopathic asthenozoospermia.
OBJECTIVES: To investigate the expression level of testis-specific calcium-binding protein CBP86-IV in normal and asthenozoospermic human sperm. METHODS: 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. RESULTS: 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. CONCLUSIONS: The stable and reliable change of TSCBP86-IV may be taken as a new molecular marker for clinical diagnosis of idiopathic asthenozoospermia.
Authors: Young-Hwan Kim; Kula N Jha; Arabinda Mandal; Geeta Vanage; Erin Farris; Phillip L Snow; Ken Klotz; Soren Naaby-Hansen; Charles J Flickinger; John C Herr Journal: Dev Biol Date: 2005-10-01 Impact factor: 3.582