Introduction of mouse C epsilon genes into Cos-7 cells and fertilized mouse eggs.
A fragment of the cloned gene for the mouse C epsilon chain, coding for the first, second, third, and fourth domains, has been coupled to the SV40 promotor region (pSV2-mC epsilon). About 50 copies of pSV2-mC epsilon or its PvuII-EcoRI fragments were introduced into Cos-7 cells. Expression of PvuII-EcoRI fragments of pSV2-mC epsilon was observed in about 50% of the Cos-7 cells by indirect fluorescence staining. However, no expression of circular pSV2-mC epsilon was observed. About 200 copies of linearized pSV2-mC epsilon with EcoRI were introduced into fertilized mouse eggs. Two of 78 mice born from these eggs had integrated mouse C epsilon genes. Mouse C epsilon gene was shown to be integrated in a tandem array and as intact structures without undergoing gross deletions or rearrangements, judged from the Southern blotting patterns from several restriction enzymes. The first transgenic mouse was mated to a normal male to examine whether mouse C epsilon gene were stably transmitted to progeny. Among 5 mice to which the C epsilon gene had been transmitted, one deleted 5 copies of this gene and another deleted one junction fragment, thus demonstrating relatively unstable transmission. No C epsilon mRNA was detected in the liver, kidney, brain, lung, skeletal muscle, heart, testis, or spleen of a transgenic mouse.
Yamamura
K
K
Miki
T
T
Suzuki
N
N
Ebihara
T
T
Kawai
K
K
Kumahara
Y
Y
Honjo
T
T
D000818:Animals; D002460:Cell Line; D003001:Cloning, Molecular; D004247:DNA / Q000235:genetics; D004270:DNA, Circular / Q000235:genetics; D004274:DNA, Recombinant; D005260:Female; D005818:Genetic Engineering; D007136:Immunoglobulins / Q000235:genetics; D008297:Male; D051379:Mice; D009693:Nucleic Acid Hybridization; D012333:RNA, Messenger / Q000032:analysis; D011995:Recombination, Genetic; D013539:Simian virus 40 / Q000235:genetics; D014158:Transcription, Genetic; D015053:Zygote
D004270:DNA, Circular; D004274:DNA, Recombinant; D007136:Immunoglobulins; D012333:RNA, Messenger; D004247:DNA