Quantitative analysis of the human alpha-skeletal actin gene in transgenic mice.
Three aspects of the regulation of the human alpha-skeletal actin gene are examined in this study by quantitative analysis of transgenic tissues: level of expression, tissue specificity, and developmental regulation. Previous in vitro and in vivo studies analyzing the 5' end of the gene have indicated that regulation of tissue-specific expression is promoter based. Transgenic mice were produced carrying either a 9.5-kilobase pair (kb) human alpha-skeletal actin gene fragment or a deletion construct with 2.2-kb of 5' sequences of human alpha-skeletal actin linked to the chloramphenicol acetyltransferase reporter gene. We found that the 9.5-kb transgene was capable of expression in adult skeletal muscle at a level equivalent to that of the endogenous gene in a non-transgenic mouse. The deletion construct was also capable of high-level expression. Both transgenes were expressed in a striated muscle-specific manner and were correctly regulated during development. We conclude that these three parameters of regulation of the human alpha-skeletal actin gene are mediated by sequences within the region -2000 to +239 of the promoter.
Brennan
K J
KJ
Muscle Development Unit, Children's Medical Research Institute, Wentworthville, New South Wales, Australia.
Hardeman
E C
EC
D000199:Actins / Q000096:biosynthesis / Q000235:genetics*; D000375:Aging / Q000502:physiology; D000818:Animals; D015500:Chloramphenicol O-Acetyltransferase / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism; D015342:DNA Probes; D006321:Heart / Q000502:physiology; D006801:Humans; D051379:Mice; D008822:Mice, Transgenic; D009130:Muscle, Smooth / Q000502:physiology; D009132:Muscles / Q000502:physiology*; D009928:Organ Specificity; D012313:RNA / Q000032:analysis; D012333:RNA, Messenger / Q000032:analysis* / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000378:metabolism; D015183:Restriction Mapping; D017384:Sequence Deletion
D000199:Actins; D015342:DNA Probes; D012333:RNA, Messenger; D011993:Recombinant Fusion Proteins; D012313:RNA; D015500:Chloramphenicol O-Acetyltransferase