Material & MethodsPatients and Semen Donors1997 1998 139 subfertility infertility
Conventional PCR AssaysLeukocytes DNA PCR amplification Deletions loci : sY81, sY84, sY182, sY94, sY102, sY117, sY143, sY147 sY152, sY153, and sY157.
FISH sequenced DAZ cosmids FISH probes . DAZ 5 Cosmid 18E8 (Genbank AC010089) exons 2 11 Cosmos 63C9 (Genbank AC000021) exons 2 11 35 kb downstream DAZ 3 Cosmid 46A6 (Genbank AC000022) 18E8 46A6 combination probes DAZ gene 5 3 detection 18E8 63C9 combination of probes 5 single clusters DAZ gene detection Fiber FISH .Probe biotin digoxigenin labeling, target DNA avidin antidigoxigenin antibodies fluorochromes Cy3 (red) fluorescein (green) hybridization detectecting.
Discussion1. Y DAZ gene 3 autosomal DAZL transposition . Intragenic amplification, exon pruning duplication Y head-to-head manner 4 .2. sY581 marker exons 2 6 DAZ gene 10.8-kb segment detect probes . region RNA recognition motif DAZ gene clusters motif 7 .: DAZ1 , DAZ4 , DAZ2 DAZ3 .3. partial deletions DAZ gene clusters . AZFb deletion cluster 1 Y deletion . cluster 1 AZFb cluster cluster 2 cluster . AZFb deletion AZFc deletion . breakpoints overlap , intervals
4. 4 . DAZ . 4 DAZ RNA recognition motifs . .
5. clusters . less severe phenotype cluster DAZ . cluster1 cluster2 .
6. DAZ gene cluster 6 3 sperm 5 X 106 cells/mL . deletions deletion .
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