Shen Qingmiao, Li Junying, Bao Haigang, Wu Changxin
National Engineering Laboratory for Animal Breeding, Beijing Key Laboratory of Animal Genetic Improvement, College of Animal Science and Technology, China Agricultural University, Beijing 100193, China.
Animals (Basel). 2023 Mar 18;13(6):1091. doi: 10.3390/ani13061091.
Sex-linked phenotypes of late feathering (LF) and early feathering (EF) are controlled by a pair of alleles and . Autosexing based on the feathering rate is widely used in poultry production. It is reported that a tandem duplication of 176,324 base pairs linked to the locus is responsible for LF expression and could be used as a molecular marker to detect LF chicken. So far, there is no genotyping method that can accurately and stably identify the LF homozygote and heterozygote in all chicken breeds. In the present study, a multiplex PCR test was developed to identify EF, LF homozygote, and heterozygote according to electrophoretic bands and the relative height of the peaks by Sanger sequencing. We tested 413 chickens of six native Chinese breeds with this method. The identification was consistent with the sex and phenotype records of the chickens. Band density analysis was performed, and the results supported our genotyping using the new assay. In order to further verify the accuracy of this test in distinguishing homozygote and heterozygote males, 152 LF males were mated with EF females, and the results of the offspring's phenotypes were consistent with our expectations. Our results support tandem duplication as molecular markers of LF, and this new test is applicable to all LF chickens associated with tandem duplication.
迟羽（LF）和早羽（EF）的伴性表型由一对等位基因控制。基于羽速的自别雌雄在家禽生产中被广泛应用。据报道，与该基因座连锁的176,324个碱基对的串联重复导致迟羽表达，可作为检测迟羽鸡的分子标记。到目前为止，还没有一种基因分型方法能够在所有鸡品种中准确、稳定地鉴定迟羽纯合子和杂合子。在本研究中，开发了一种多重PCR检测方法，根据电泳条带和桑格测序峰的相对高度来鉴定早羽、迟羽纯合子和杂合子。我们用这种方法检测了6个中国本土品种的413只鸡。鉴定结果与鸡的性别和表型记录一致。进行了条带密度分析，结果支持我们使用新检测方法进行的基因分型。为了进一步验证该检测方法在区分纯合子和杂合子雄性方面的准确性，将152只迟羽雄性与早羽雌性进行交配，后代的表型结果与我们的预期一致。我们的结果支持串联重复作为迟羽的分子标记，并且这种新检测方法适用于所有与串联重复相关的迟羽鸡。
