| Project/Area Number |
18580291
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Basic veterinary science/Basic zootechnical science
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| Research Institution | Saga University |
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Principal Investigator |
WADA Yasuhiko Saga University, Faculty of Agiculture, Professor (40325585)
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| Co-Investigator(Kenkyū-buntansha) |
NISHIBORI Masahide Hiroshima University, Graduate School of Biosphere Science, Associate Professor (80237718)
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| Project Period (FY) |
2006 – 2007
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| Project Status |
Completed (Fiscal Year 2007)
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| Budget Amount *help |
¥3,560,000 (Direct Cost: ¥3,200,000、Indirect Cost: ¥360,000)
Fiscal Year 2007: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
Fiscal Year 2006: ¥2,000,000 (Direct Cost: ¥2,000,000)
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| Keywords | Silkie chicken / genome analysis / prolactin / Egg production ratio / DNA marker / PCR-RFLP / SNP / Japanese native chicken / ゲノム / メラニン色素生成 / 就巣性 / チロシナーゼ / ゲノミックサブトラクション / TRP1 / TRP2 |
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| Research Abstract |
Silkie chicken (Gallus gallus) have played an important role in traditional Chinese Medicine, however the genes associated with the specific appearance and deposition of the melanin pigment are unknown. The three genomic subtraction libraries were constructed using PCR subtraction method between a Silkie chicken and a White Leghorn chicken. We sequenced 178 clones of these libraries and found the 86 unique clones. The primer pairs for PCR-RFLP system were examined for the Silkie chicken and the White Leghorn chicken. The polymorphism between Silkie chicken and White Leghorn was found in the thirteen primer pairs. The 5% level significant differences of allele frequency between Silkie chicken and the other breeds were found in two PCR-RFLP primer pairs.
The 24-bp insertion of the promoter region of prolactin gene was studied for 53 Silkie fowls, 19 Japanese native chickens and 6 commercial chickens. Forty-six Silkie fowls have genotype Del/Del, 7 Silkie fowls have genotype In/Del, howeve
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r no Silkie fowls have genotype In/In. On the other hand, 11, 8 and 6 birds of other chickens showed genotypes Del/Del, In/Del and In/In, respectively. The chi-squared test using 2x3 table showed the significant difference (P<0.01) of the genotype frequencies between Silkie fowls and other chickens. These results show the significant difference of the genotype frequencies for the 24-bp insertion of the promoter region of prolactin gene between Silkie fowls and other chickens.
Silkie chicken (Gallus gallus) have played an important role in traditional Chinese Medicine, however broodiness is often observed and egg production ratio is very small in Silkie chicken. It is thought that prolactin gene is an essential for the start and maintain of broodiness in hens. The 24-bp insertion is found in the promoter region of prolactin gene of White Leghorn as compare with Red Jungle fowl and the absence of the 24-bp insertion would maintain normal function of the prolactin promoter. In this study, the 24-bp insertion was genotyped in the fourth generation of the Silkie population selected for egg production and the relationship between the genotype of the insertion locus and performance traits was investigated. Blood samples of 69 male Silkies and 242 female Silkies were collected form the fourth generation of Silkie population selected for egg production in Livestock Research Institute of Oita Prefectural Agriculture, Forestry and Fisheries Research Center. Genomic DNA was isolated from blood samples and the fragment of the promoter of prolactin gene was amplified using PCR. The PCR products were electrophoresed on 3.0% agarose gels for genotyping. The 216 Del/Del Silkies, 26 In/Del Silkies and no In/In Silkie were found in this study. The relationship between the genotype of the insertion locus and the performance traits was analyzed by analysis of variance, however there was no relationship for growth traits and egg production. The In/Del Silkies have higher peak of the distribution of 151d-450d egg production ratio than the De/Del Silkies. Less
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