| Summary: | 碩士 === 國立中興大學 === 畜產學系 === 89 === The purpose of this research is to amplify sex-specific DNA
fragments of Holstein cattle, Taiwan yellow cattle, black and white Taiwan water buffalo using PCR, and then to compare diversity among species combined with sequencing.
The results of RAPD-PCR amplified with OPC and OPD operon
indicated that OPD02 primer（5’-GGACCCAACC-3’）could amplify a
673-bp sex-specific band in black Taiwan water buffalo, but it does not appear in white Taiwan water buffalo. Furthermore, two sex-specific bands could be amplified by OPD15 primer（5’-CATCCGTGCT-3’）both in black and white Taiwan water buffalo. The longer one is not the same with white Taiwan water buffalo from black one, otherwise the shorter one has 94.7% similarity between black and white Taiwan water buffalo. And, there are total 20 nucelotide-variations among the shorter bands,
incoulding 3 deletion, 11 transition and 6 transversion. Except Taiwan water buffalo, Holstein cattle and Taiwan yellow cattle could not be amplified any sex-specific pattern by using both OPC and OPD serial primers.
According to the Japanese Sika deer sex-specific DNA sequence, MT1 and MT2 primer pair could amplify a sex-specific band in all of male Holstein cattle, Taiwan yellow cattle, black and white Taiwan water buffalo. And, there are some differences in nucleotides of this band between Holstein cattle and black Taiwan water buffalo.
Using BosSry1（ 5’-CCACGTCAAGCGACCCATGAAC-3’）and BosSry2（5’-TCAGCATCTGTAAGCCTTTTCCACTC-3’）primer pair consulted from Bos taurus Sry cDNA（2831 bp）and Japanese Sika deer Sry cDNA（690 bp）, a band could be amplified in all of male Holstein cattle, Taiwan yellow cattle, black and white Taiwan water buffalo. And, there are some differences in nucleotides of this band among Holstein cattle, Taiwan yellow cattle, black and white Taiwan water buffalo.
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