ABSTRACT
Four breeds of chickens (White Leghorn, Yangshan, Taihe Silkies, White Recessive Rocks) with different egg production were applied to screen potential single nucleotide polymorphisms (SNPs) related to reproduction trait in distal part region of prolactin gene based on DNA pooling and sequencing. Eight SNPs (C2402T, T-2192C, C-2161G, C-2134G, C-2062G, G-2040A, A-1944G and C-1884A) were found successfully in the fragment of 1 028 bp at the distal part of 5'flanking region. Moreover,a simple and rapid method to estimate allelic frequencies of SNPs in each breed by calculating the ratio of allele peak heights was introduced here. The accuracies of estimation for the allelic frequencies of C-2402T, C-2161G, C-1884A, C-2062G and G-2040A were verified by comparing with the results of PCR-RFLP or PCR-SSCP.
Subject(s)
Chickens/genetics , DNA/genetics , Polymorphism, Single Nucleotide , Prolactin/genetics , 5' Flanking Region , Animals , Base Sequence , Chickens/classification , DNA/chemistry , Gene Frequency , Genome , Genotype , Molecular Sequence Data , Polymerase Chain Reaction , Polymorphism, Restriction Fragment Length , Polymorphism, Single-Stranded Conformational , Quantitative Trait Loci , Sequence Analysis, DNAABSTRACT
Four chicken breeds (White Leghorn, Yangshan, Taihe Silkies, White Recessive Rocks) with different reproduction were applied to screen potential SNPs related to laying performance in the 5' flanking region, exon region and partial intron region of chicken prolactin (cPRL) gene. Totally almost 4500 bp were screened rapidly based on DNA pooling and sequencing, and thirteen single nucleotide polymorphisms (SNPs) and two indels (24 bp and 15 bp) were found, including nine SNPs and two indels in the 5' flanking region, one SNP in Exon 2, two SNPs in Exon 5 and one SNP in Intron 2 respectively. Furthermore, 5' flanking region of cPRL gene was analyzed by the website of http://motif.genome.ad.jp/. A possible Evi-1 binding site (score 93) was found in White Leghorn cPRL gene because of the 24 bp insertion, another possible C/EBPbeta binding site (score 94) was found in Yangshan cPRL gene because of the variation of C-2402T. Further studies need to be carried out to verify their effects on the expression of cPRL gene, the broodiness and laying performance of chickens.
