Identification of Duplication Genotypes of the Feathering Rate Gene in Chicken by a Multiplex PCR Following Electrophoresis and/or Sanger Sequencing
Sex-linked phenotypes of late feathering (LF) and early feathering (EF) are controlled by a pair of alleles <i>K</i> and <i>k<sup>+</sup></i>. Autosexing based on the feathering rate is widely used in poultry production. It is reported that a tandem duplication of...
| Main Authors: | , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2023-03-01
|
| Series: | Animals |
| Subjects: | |
| Online Access: | https://www.mdpi.com/2076-2615/13/6/1091 |
| _version_ | 1797613959851802624 |
|---|---|
| author | Qingmiao Shen Junying Li Haigang Bao Changxin Wu |
| author_facet | Qingmiao Shen Junying Li Haigang Bao Changxin Wu |
| author_sort | Qingmiao Shen |
| collection | DOAJ |
| description | Sex-linked phenotypes of late feathering (LF) and early feathering (EF) are controlled by a pair of alleles <i>K</i> and <i>k<sup>+</sup></i>. 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 <i>K</i> 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. |
| first_indexed | 2024-03-11T07:01:55Z |
| format | Article |
| id | doaj.art-c923123a4e894be5ac76cda5708d9643 |
| institution | Directory Open Access Journal |
| issn | 2076-2615 |
| language | English |
| last_indexed | 2024-03-11T07:01:55Z |
| publishDate | 2023-03-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Animals |
| spelling | doaj.art-c923123a4e894be5ac76cda5708d96432023-11-17T09:12:15ZengMDPI AGAnimals2076-26152023-03-01136109110.3390/ani13061091Identification of Duplication Genotypes of the Feathering Rate Gene in Chicken by a Multiplex PCR Following Electrophoresis and/or Sanger SequencingQingmiao Shen0Junying Li1Haigang Bao2Changxin Wu3National Engineering Laboratory for Animal Breeding, Beijing Key Laboratory of Animal Genetic Improvement, College of Animal Science and Technology, China Agricultural University, Beijing 100193, ChinaNational Engineering Laboratory for Animal Breeding, Beijing Key Laboratory of Animal Genetic Improvement, College of Animal Science and Technology, China Agricultural University, Beijing 100193, ChinaNational Engineering Laboratory for Animal Breeding, Beijing Key Laboratory of Animal Genetic Improvement, College of Animal Science and Technology, China Agricultural University, Beijing 100193, ChinaNational Engineering Laboratory for Animal Breeding, Beijing Key Laboratory of Animal Genetic Improvement, College of Animal Science and Technology, China Agricultural University, Beijing 100193, ChinaSex-linked phenotypes of late feathering (LF) and early feathering (EF) are controlled by a pair of alleles <i>K</i> and <i>k<sup>+</sup></i>. 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 <i>K</i> 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.https://www.mdpi.com/2076-2615/13/6/1091chickenfeathering ratetandem duplicationmultiplex PCR |
| spellingShingle | Qingmiao Shen Junying Li Haigang Bao Changxin Wu Identification of Duplication Genotypes of the Feathering Rate Gene in Chicken by a Multiplex PCR Following Electrophoresis and/or Sanger Sequencing Animals chicken feathering rate tandem duplication multiplex PCR |
| title | Identification of Duplication Genotypes of the Feathering Rate Gene in Chicken by a Multiplex PCR Following Electrophoresis and/or Sanger Sequencing |
| title_full | Identification of Duplication Genotypes of the Feathering Rate Gene in Chicken by a Multiplex PCR Following Electrophoresis and/or Sanger Sequencing |
| title_fullStr | Identification of Duplication Genotypes of the Feathering Rate Gene in Chicken by a Multiplex PCR Following Electrophoresis and/or Sanger Sequencing |
| title_full_unstemmed | Identification of Duplication Genotypes of the Feathering Rate Gene in Chicken by a Multiplex PCR Following Electrophoresis and/or Sanger Sequencing |
| title_short | Identification of Duplication Genotypes of the Feathering Rate Gene in Chicken by a Multiplex PCR Following Electrophoresis and/or Sanger Sequencing |
| title_sort | identification of duplication genotypes of the feathering rate gene in chicken by a multiplex pcr following electrophoresis and or sanger sequencing |
| topic | chicken feathering rate tandem duplication multiplex PCR |
| url | https://www.mdpi.com/2076-2615/13/6/1091 |
| work_keys_str_mv | AT qingmiaoshen identificationofduplicationgenotypesofthefeatheringrategeneinchickenbyamultiplexpcrfollowingelectrophoresisandorsangersequencing AT junyingli identificationofduplicationgenotypesofthefeatheringrategeneinchickenbyamultiplexpcrfollowingelectrophoresisandorsangersequencing AT haigangbao identificationofduplicationgenotypesofthefeatheringrategeneinchickenbyamultiplexpcrfollowingelectrophoresisandorsangersequencing AT changxinwu identificationofduplicationgenotypesofthefeatheringrategeneinchickenbyamultiplexpcrfollowingelectrophoresisandorsangersequencing |