Cloning, Structure and Regulation of the sp56 Gene

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9509863 Bleil The primary interest of Dr. Bleil's laboratory is analysis of mammalian fertilization at the molecular level, combining techniques of cell biology, biochemistry and molecular genetics. He has identified a mouse sperm protein, a lectin called sp56, which has many properties expected of the protein responsible for sperm recognition of the mouse egg's zona pellucida. He has recently isolated and cloned the cDNA encoding sp56. Amino acid sequence derived from this cDNA indicates that: 1) the protein is translated on the endoplasmic reticulum, 2) has no transmembrane domain, and 3) is a member of a superfamily of extracellular protein receptors. Probes directed against sp56 and its mRNA reveal that transcription of the sp56 gene and translation of its mRNA occur exclusively in spermatogenic cells of the mouse. The long term goals to be addressed by studies described here are: 1) to genetically prove or disprove the hypothesis that sp56 is the sperm protein which recognizes the mouse egg zona pellucida, mediating sperm binding to the egg's zona pellucida, and 2) to determine the molecular structure of the sp56/ZP3 complex. In order to attain these goals, the gene encoding sp56 must be identified and isolated. The specific aims of the proposed study are as follows: 1) To use cDNA encoding sp56, currently available in this laboratory, to screen a mouse genomic library. Genomic DNA clones, including sequence upstream of the translated region, will be purified and sequenced; 2) To analyze the sequence of the sp56 gene in order to identify regulatory sequences which might encode specific expression in testes. Comparison with other, previously identified genes which are expressed in testes may reveal common regulatory sequences. ***