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  • Title: Expression of a gene encoding a specific human sperm protein in Chinese hamster ovarian cells.
    Author: Miao SY, Liu QY, Wang LF, Zhao M, Yan YC, Koide SS.
    Journal: Arch Androl; 1992; 28(2):91-7. PubMed ID: 1520040.
    Abstract:
    The cDNA (RSD-2) encoding a human sperm protein (YWK-II) was isolated from a rat testis lambda gt11 expression library. Its nucleotide sequence was determined. The deduced polypeptide showed high identity with the transmembrane-cytoplasmic domains of A4 amyloid precursor protein of Alzheimer's disease. The RSD-2 was inserted into the EcoRI site of the pSV2-EP vector to construct the pSVRS-2 vector. Chinese hamster ovarian (CHO-dhfr) cells were cotransformed with pSV2-neo and pSVRS-2. mRNA and chromosomal DNA prepared from the transformed cells interacted with [32P]RSD-2 as probe by dot hybridization. The production of the YWK-II protein was determined by staining with the YWK-II mAb by an indirect immunofluorescence technique. There was marked staining of the cytoplasm. The RSD-2 cDNA encoding the YWK-II sperm was expressed in the transformed CHO cells. The pSV2-EP vector and the CHO cell expression system can be utilized to produce sperm proteins for antifertility studies.
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