title: A novel minisequencing single-nucleotide polymorphism marker of the lysozyme gene detects high hatchability of Tsaiya ducks (Anas platyrhynchos).
abstract: Lysozyme, one of the major albumen antimicrobials, can break down the polysaccharide walls of a broad spectrum of bacteria. This study presents a novel lysozyme marker of high hatchability in the form of minisequencing single-nucleotide polymorphisms (SNPs). Recently, lysozyme was identified by complementary DNA microarray analysis as one of several differentially expressed genes noted to influence hatchability and recognized as a marker candidate for animal marker-assisted selection. Higher levels (P < 0.05) of lysozyme mRNA (via real-time polymerase chain reaction analysis) and protein (in Western blotting results) were found to be associated with a high-hatchability phenotype. In the preliminary sequence analysis of this study, TsLy1-1 and TsLy1-2 primer pairs, designed according to the lysozyme sequence, were used to amplify small-scale genomic DNA samples from animals in two extreme groups of hatchability. Sequence analysis of the amplified 763-bp DNA products clearly showed that AA and GG genotypes of SNP g.390A > G were from the ducks of the low- and high-hatchability groups, respectively. The SNP g.390A > G also created a new specificity protein 1 transcription factor binding site in the lysozyme gene. Primer pairs of TsLy2-1 and TsLy2-2 then probed the amplified 763-bp DNA products to produce a shorter fragment for easier minisequencing analysis to divide 114 ducks into GG, GA, and AA genotypes. The GG ducks had the highest hatchability, representing that a new lysozyme SNP marker of good hatchability performance can be used for the purpose of marker-assisted selection in Tsaiya ducks.
authors.affiliation: Department of Biotechnology, MingDao University, ChangHua, Taiwan. Electronic address: hlhuang@mdu.edu.tw.
authors.forename: Hsiu-Lin
authors.initials: HL
authors.lastname: Huang
authors.affiliation: Livestock Research Institute, Council of Agriculture, Hsin-Hua, Tainan, Taiwan.
authors.forename: Yu-Shin
authors.initials: YS
authors.lastname: Cheng
mesh_terms: D000818:Animals; D001483:Base Sequence; D004247:DNA / Q000737:chemistry; D004372:Ducks / Q000235:genetics*; D015870:Gene Expression; D005819:Genetic Markers / Q000235:genetics*; D005838:Genotype; D009113:Muramidase / Q000032:analysis / Q000235:genetics*; D010063:Ovum / Q000502:physiology*; D020641:Polymorphism, Single Nucleotide / Q000235:genetics*; D012333:RNA, Messenger / Q000032:analysis; D060888:Real-Time Polymerase Chain Reaction; D012098:Reproduction / Q000235:genetics*; D012641:Selection, Genetic; D017422:Sequence Analysis, DNA / Q000662:veterinary
chemical_list: D005819:Genetic Markers; D012333:RNA, Messenger; D004247:DNA; D009113:Muramidase
keywords: Antimicrobial; Hatchability; Lysozyme gene; Marker-assisted selection; Tsaiya duck