Detection of chicken dominant white gene <I>PMEL17</I> mutation site by a pooling method

Abstract

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.