Mitochondrial DNA detection and copy number determination in the spermatozoa of the sea urchin Arbacia lixula.
The Polymerase Chain reaction technique has been used in order to detect and amplify a specific region of mtDNA, in a total DNA preparation extracted from the sperm of the sea urchin Arbacia lixula. The amplified fragment is the D-loop region which hybridizes with the homologous region extracted from the egg mtDNA. The results demonstrate that mtDNA is present in sperm cell, and, since the replication origin is present it is potentially able to replicate in the zygote. Furthermore, the technique used allowed us to estimate mtDNA copy number in sea urchin sperm, which has never been done before. Our results are that sea urchin sperm cell contains between 4 and 28 mtDNA molecules.
De Giorgi
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Dipartimento di Biochimica e Biologia Molecolare, Università di Bari, Via, Italy.
D'Alessandro
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Saccone
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D000818:Animals; D001482:Base Composition; D001483:Base Sequence; D004272:DNA, Mitochondrial / Q000032:analysis* / Q000235:genetics; D005260:Female; D008297:Male; D008969:Molecular Sequence Data; D009838:Oligodeoxyribonucleotides; D010063:Ovum / Q000737:chemistry / Q000502:physiology; D016133:Polymerase Chain Reaction; D012617:Sea Urchins; D012689:Sequence Homology, Nucleic Acid; D013094:Spermatozoa / Q000737:chemistry / Q000502:physiology*
D004272:DNA, Mitochondrial; D009838:Oligodeoxyribonucleotides