Coexpression of epsilon, G gamma and A gamma globin mRNA in embryonic red blood cells from a single copy beta-YAC transgenic mouse.
We utilized reverse transcription polymerase chain reaction (RT-PCR) to amplify epsilon, G gamma and A gamma globin cDNAs from single red blood cells isolated from a day-10 transgenic fetus harboring a single copy of the human beta-YAC. A detailed structural analysis of the beta-YAC showed a single copy of each beta-like globin gene is present and linked to the locus control region (LCR). RNase protection analysis of RNA isolated from erythroid tissues from day-8 to day-16 of development and the adult stage showed proper developmental switching of the beta-like globin gene expression. Using epsilon / gamma and G gamma / A gamma primer sets in separate RT-PCR reactions on RNA from single day-10 red blood cells we observed an intercellular variation in the epsilon and gamma RT-PCR products that may be indicative of a change in the LCR preference from the epsilon gene promoter to the gamma gene promoter during switching. We also found that the majority of the red blood cells examined contain all three globin mRNA species. These observations suggest that either the LCR is capable of interacting simultaneously with more than one globin gene promoter or alternatively, the LCR may interact with only one promoter at any given time, but its interaction oscillates between promoters (flip-flop mechanism) resulting in expression of more than one gene from a single beta-globin locus.
Furukawa
T
T
Department of Medicine, University of Washington, Seattle 98195, USA.
Navas
P A
PA
Josephson
B M
BM
Peterson
K R
KR
Papayannopoulou
T
T
Stamatoyannopoulos
G
G
D000818:Animals; D001483:Base Sequence; D004912:Erythrocytes / Q000378:metabolism*; D005260:Female; D005312:Fetal Blood / Q000378:metabolism*; D018507:Gene Expression Regulation, Developmental; D005914:Globins / Q000096:biosynthesis* / Q000235:genetics; D051379:Mice; D008822:Mice, Transgenic; D008969:Molecular Sequence Data; D011247:Pregnancy; D011401:Promoter Regions, Genetic / Q000235:genetics; D012333:RNA, Messenger / Q000096:biosynthesis*
D012333:RNA, Messenger; D005914:Globins