Expression of a gene encoding a specific human sperm protein in Chinese hamster ovarian cells

Arch Androl. 1992 Mar-Apr;28(2):91-7. doi: 10.3109/01485019208987685.

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amyloid beta-Protein Precursor*
  • Animals
  • Antibodies, Monoclonal / biosynthesis*
  • Base Sequence
  • Cricetinae
  • DNA / analysis
  • Female
  • Immunoblotting
  • Membrane Proteins / biosynthesis*
  • Membrane Proteins / genetics
  • Molecular Sequence Data
  • Nerve Tissue Proteins*
  • Ovary / metabolism*
  • Plasmids
  • RNA, Messenger / analysis
  • Transformation, Genetic

Substances

  • APLP1 protein, human
  • Amyloid beta-Protein Precursor
  • Antibodies, Monoclonal
  • Aplp2 protein, rat
  • Membrane Proteins
  • Nerve Tissue Proteins
  • RNA, Messenger
  • DNA