Reliable sex identification of dogs by modified PCR/RFLP analysis.
To find definitive RFLP sites for canine sex determination, DNA segments corresponding to parts of the canine ZFX and ZFY genes were amplified by PCR and were directly sequenced. According to the newly defined sequence data, the combination of Haelll and Cfr13I sites was found to be useful for not only identifying the sex of the canine DNA samples but also distinguishing them from the human DNA. Conveniently, these two enzymes worked simultaneously in the same single buffer. The double-digestion of the ZFX/ZFY PCR products with HaeIII and Cfr13I showed banding patterns unique to males and females in Canis familialis. This PCR/RFLP method was confirmed to be applicable to various breeds of dog.
Murakami
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M
Laboratories of Molecular Biology, Azabu University School of Veterinary Medicine, Sagamihara, Kanagawa, Japan.
Fujise
H
H
Lee
Y S
YS
Matsuba
C
C
Fujitani
H
H
D000818:Animals; D001483:Base Sequence; D004247:DNA / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D004268:DNA-Binding Proteins / Q000235:genetics*; D015252:Deoxyribonucleases, Type II Site-Specific / Q000737:chemistry; D004285:Dogs / Q000235:genetics* / Q000502:physiology; D005260:Female; D051741:Kruppel-Like Transcription Factors; D008297:Male; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000662:veterinary; D012150:Polymorphism, Restriction Fragment Length; D012689:Sequence Homology, Nucleic Acid; D012730:Sex Chromosomes / Q000737:chemistry / Q000235:genetics; D012732:Sex Determination Analysis / Q000379:methods / Q000662:veterinary*; D014157:Transcription Factors
D004268:DNA-Binding Proteins; D051741:Kruppel-Like Transcription Factors; D014157:Transcription Factors; C062076:zinc finger protein, X-linked; D004247:DNA; D015252:Deoxyribonucleases, Type II Site-Specific; C024403:GGCC-specific type II deoxyribonucleases; C032262:GGNCC-specific type II deoxyribonucleases