Tracking H-2 alleles in transgenic mice by RFLP and heteroduplex analysis.
Transgenic mice provide valuable tools for biological research including many areas of immunology. In studies involving the major histocompatibility complex (MHC), it is often necessary to place the desired transgene in a specific H-2 (the murine MHC) environment. In this regard, the strains commonly used for the production of transgenic mice also carry well characterized H-2 alleles and provide an appropriate genetic background for MHC related experiments. In this study, a highly polymorphic microsatellite of tetranucleotide repeats from the second intron of the class II Eb gene within the H-2 complex was used in order to identify the corresponding alleles. The relevant H-2 allele(s) along with the transgene were then tracked throughout the production of a chicken ovalbumin-specific transgenic strain. The technique involved PCR-amplification of a DNA sequence encompassing the H-2 specific microsatellite followed by RFLP and heteroduplex analyses. This approach is likely to find wide application in the background checking of transgenic mice, especially in immunological research requiring a defined H-2 background.
Shanmugam
V
V
Department of Pathology, Emory University School of Medicine, Atlanta, GA 30322, USA.
Haines
D
D
Lake
J P
JP
Saha
B K
BK
D000483:Alleles; D000818:Animals; D001483:Base Sequence; D005819:Genetic Markers; D006183:H-2 Antigens / Q000032:analysis / Q000235:genetics*; D051379:Mice; D008822:Mice, Transgenic; D018895:Microsatellite Repeats; D008969:Molecular Sequence Data; D009692:Nucleic Acid Heteroduplexes / Q000032:analysis*; D009693:Nucleic Acid Hybridization; D016133:Polymerase Chain Reaction; D012150:Polymorphism, Restriction Fragment Length; D016415:Sequence Alignment; D012689:Sequence Homology, Nucleic Acid
D005819:Genetic Markers; D006183:H-2 Antigens; D009692:Nucleic Acid Heteroduplexes