Transgene insertion pattern analysis using genomic walking in a transgenic mouse line.
A transgene mapping technique (Noguchi et al., Exp. Anim. 53:103-111, 2004) is described that can be used to analyze transgene integration patterns in transgenic mice. The technique was used to reveal that a transgenic mouse line (GM1-sy#116) harbored inverted and direct tandem repeats of both intact and partial pCAGGS-based transgenes in the G2 region of chromosome 1. This complicated concatenation of transgenes may have been caused by simple end-joining of DNA constructs fragmented by exposure to UV transillumination during gel-purification, and by nuclease digestion inside zygote pronuclei. The results suggest that care should be taken to avoid unwanted fragmentation during the preparation of vector constructs.
Suzuki
Osamu
O
Department of Veterinary Science, National Institute of Infectious Diseases, Toyama, Tokyo, Japan.
Hata
Tomoko
T
Takekawa
Naho
N
Koura
Minako
M
Takano
Kaoru
K
Yamamoto
Yoshie
Y
Noguchi
Yoko
Y
Uchio-Yamada
Kozue
K
Matsuda
Junichiro
J
D000818:Animals; D016386:Chromosome Walking; D015698:Genomic Library; D005838:Genotype; D051379:Mice; D008822:Mice, Transgenic / Q000235:genetics*; D011995:Recombination, Genetic; D019076:Transgenes / Q000235:genetics*