New Screening Test for Male Germline Chimeric Chickens by Polymerase Chain Reaction Using Shingle Nucleotide Polymorphism Detection Primers

Abstract

Since it is impossible to distinguish germline chimeric chickens by appearance only, a progeny test is required for analyzing germline chimerism. To reduce the labor, time and expense involved in such test, a new screening test for male putative germline chimeric chickens by polymerase chain reaction (PCR) was developed that uses single nucleotide polymorphism (SNP) detection primers. Putative germline chimeric chickens were produced by the transfer of stage X blastodermal cells or circulating primordial germ cells from Barred Plymouth Rocks (BPR) to White Leghorns (WL). For screening prior to the progeny test, DNA was extracted from the semen and used for PCR analysis, using SNP detection primers at position 686 in the chicken mitochondrial DNA D-loop region (DNA database, AB091008). In this study, the type of SNP in all BPR at position 686 was fixed as base G, and as base A in all WL. When the PCR product was typed as both donor (base G)- and recipient (base A)-derived, the male was determined as the germline chimera. The male and female putative germline chimeric chickens produced in this study were given the progeny test. The results of the male screening test showed good agreement with the progeny test for detecting germline chimeric chickens.

Keywords

• chimeric chicken
• germline
• mitochondrial dna d-loop region
• single nucleotide polymorphism
• sperm