Detection of chicken dominant white gene PMEL17 mutation site by a pooling method
Received date: 2009-08-13
Revised date: 2009-12-10
Online published: 2010-01-15
Dominant white locus is one of the major loci affecting feather color in the domestic chicken and its dominant allele I can inhibit the synthesis of the melanin. Therefore, the homozygotes (I/I) or heterozygotes (I/i) show a white phenotype. It has been confirmed that the Dominant white locus encodes PMEL17 protein which is a specific protein and plays a key role in the development of melanocytes, thus PMEL17 gene is identified as a positional candidate gene for the dominant white phenotype in chicken. In our present study, we created an economic and efficient pooling method for detecting PMEL17 mutations in large populations, known as PCR product pooling method, and the steps are as follows: firstly, PMEL17 segments containing the mutation site from individual genomic DNA samples were amplified by PCR; secondly, 10 PCR products were mixed in a pool, and then the pooled PCR samples were separated on non-denatured PAGE gels; and finally, the mutation profile of PMEL17 in certain populations were analyzed. In addition, a comparative study between the genomic DNA pooling and the PCR product pooling method was performed, and the mutation of PMEL17 was also ana-lyzed in our experimental population. In conclusion, the PCR product pooling method proved to be appropriate power to test gene mutations.
Key words： dominant white; PMEL17; polling method; electrophoresis
LIU Wen-Bo, CHEN Sai-Rui, ZHENG Jiang-Xia, XU Gui-Yun, LI Dun-Yang, QU Lu-Jiang, YANG Ning . Detection of chicken dominant white gene PMEL17 mutation site by a pooling method[J]. Hereditas(Beijing), 2010 , 32(2) : 148 -152 . DOI: 10.3724/SP.J.1005.2010.00148
[1] Smyth JR. Genetics of Plumage Skin and Pigmentation in Chickens. In: Crawford RD. Poultry Breeding and Genetics. Elsevier Science, NY, 1990.
[2] Kerje S, Sharma P, Gunnarsson U, Kim H, Bagchi S, Fredriksson R, Schütz K, Jensen P, von Heijne G, Okimoto R, Andersson L. The Dominant white, Dun and Smoky color variants in chicken are associated with inser-tion/deletion polymorphisms in the PMEL17 gene. Genetics, 2004, 168(3): 1507–1518.
[3] Berson JF, Harper DC, Tenza D, Raposo G, Marks MS. Pmel17 initiates premelanosome morphogenesis within multivesicular bodies. Mol Biol Cell, 2001, 12(11): 3451–3464.
[4] Darvasi A, Scoller M. Selective DNA pooling for determina- tion of linkage between a molecular marker and a quantitative trait locus. Genetics, 1994, 138(4): 1365–1373.
[5] 童大跃, 伍新尧. 混合DNA样品池扩增法及其应用. 生物技术通讯, 1999, 10(3): 213–216.
[6] Lipkin E, Fulton J, Cheng H, Yonash N, Soller M. Quan-titative trait locus mapping in chickens by selective DNA pooling with dinucleotide microsatellite markers by using purified DNA and fresh or frozen red blood cells as ap-plied to marker-assisted selection. Poult Sci, 2002, 81(3): 283–292.
[7] Qu LJ, Li XY, Wu GQ, Yang N. Efficient and sensitive method of DNA silver staining in polyacrylamide gels. Electrophoresis, 2005, 26(1): 99–101
[8] Chang CM, Coville JL, Coquerelle G, Gourichon D, Oul-mouden A, Tixier-Boichard M. Complete association be-tween a retroviral insertion in the tyrosinase gene and the recessive white mutation in chickens. BMC Genomics, 2006, 7: 19.
[9] Gunnarsson U, Hellström AR, Tixier-Boichard M, Min-vielle F, Bed'hom B, Ito S, Jensen P, Rattink A, Vereijken A, Andersson L. Mutations in SLC45A2 cause plumage color variation in chicken and Japanese quail. Genetics, 2007, 175(2): 867–877.
[10] Fan E, Levin DB, Glickman BW, Logan DM. Limitations in the use of SSCP analysis. Mutat Res, 1993, 88(1): 85–92.
[11] Han YC, Teng CZ, Hu ZL, Song YC. An optimal method of DNA silver staining in polyacrylamide gels. Electro-phoresis, 2008, 29(6): 1355–1358.
[12] Sham P, Bader JS, Craig I, O'Donovan M, Owen M. DNA Pooling: a tool for large-scale association studies. Nat Rev Genet, 2002, 3(11): 862–871.
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