Expression of a gene encoding a specific human sperm protein in Chinese hamster ovarian cells.
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.
Miao
S Y
SY
Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences, Beijing.
Liu
Q Y
QY
Wang
L F
LF
Zhao
M
M
Yan
Y C
YC
Koide
S S
SS
D016564:Amyloid beta-Protein Precursor; D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis*; D001483:Base Sequence; D006224:Cricetinae; D004247:DNA / Q000032:analysis; D005260:Female; D015151:Immunoblotting; D008565:Membrane Proteins / Q000096:biosynthesis* / Q000235:genetics; D008969:Molecular Sequence Data; D009419:Nerve Tissue Proteins; D010053:Ovary / Q000378:metabolism*; D010957:Plasmids; D012333:RNA, Messenger / Q000032:analysis; D014170:Transformation, Genetic
C489697:APLP1 protein, human; D016564:Amyloid beta-Protein Precursor; D000911:Antibodies, Monoclonal; C498283:Aplp2 protein, rat; D008565:Membrane Proteins; D009419:Nerve Tissue Proteins; D012333:RNA, Messenger; D004247:DNA