THE STUDY OF SPERM PROTEIN sp32/OY-TES-1 mRNA AND PROTEIN EXPRESSION IN HUMAN NORMAL TISSUES

Acta Anatomica Sinica ›› 2007, Vol. 38 ›› Issue (6) : 675-680.

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Acta Anatomica Sinica ›› 2007, Vol. 38 ›› Issue (6) : 675-680.
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THE STUDY OF SPERM PROTEIN sp32/OY-TES-1 mRNA AND PROTEIN EXPRESSION IN HUMAN NORMAL TISSUES

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Abstract

Objective To investigate whether the expression of sp32/OY-TES-1 mRNA is various in human normal tissues, study the expression of its protein, and provide more information about sp32/OY-TES-1 expressive profile. Methods Extraction of total RNA from 40 cases of human normal tissues included 19 different types.cDNA was synthesized by reverse transcriptase and tested by conventional polymerase chain reaction (PCR). With the construction of sp32/OY-TES-1 and HPRT plasmid for preparation of standard products, standard curves and the protocol of quantitative real-time PCR (TaqMan method) were generated, and reaction system was optimized. The expressions of sp32/OY-TES-1 and HPRT were tested respectively; OY-TES-1/HPRT was used to evaluate the expression level of sp32/OY-TES-1 mRNA. Sp32/OY-TES-1 protein that expressed in normal tissues was tested by immunohistochemistry staining with polyclonal antibody against OY-TES-1 protein. Results 1.The expressive frequency of sp32/OY-TES-1 in a panel of normal tissues was 72.50% (29/40); 2.The value of OY-TES-1/HPRT was used for the quantification of sp32/OY-TES-1 mRNA expression, which was between 0.000 1 and 0.459 0 in human nontestis normal tissues and showed in the range of logarithmic normal distribution. 3.The relative quantitative expression of sp32/OY-TES-1 mRNA in human testis tissue was 44.9, which was 91-1 000 times higher than that in other normal tissues tested. No positive target protein reaction was found in testis, colon, liver, skeletal muscle, stomach, lens, or leukocyte except in the sperm and the renal tubule of kidney. Conclusion With an extensive expression in human normal tissues, the expression level of sp32/OY-TES-1 mRNA was diverse. Of a panel of normal tissues tested, testis tissue was found wih the highest expression,

Key words

Cancer-testis antigen / Sperm protein / Gene expression / Quantitative realtime PCR / Immunohistochemistry / Human

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THE STUDY OF SPERM PROTEIN sp32/OY-TES-1 mRNA AND PROTEIN EXPRESSION IN HUMAN NORMAL TISSUES[J]. Acta Anatomica Sinica. 2007, 38(6): 675-680

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