Expression of the human Dp 71 (apo-dystrophin-1) gene from a 760-kb DMD-YAC transferred to mouse cells.
A 760-kb YAC was constructed by homologous recombination in yeast, containing the genes located in the distal portion of the DMD gene. The YAC was introduced in mouse LA-9 cells by PEG-mediated cell fusion. One transformant accommodated an intact DMD-YAC, i.e. a full copy of the DMD internal Dp 116, Dp 71 and Dp 40 genes (apo-dystrophin-2, -1 and -3, respectively). We have studied the expression of the various gene products derived from the introduced DMD-YAC. RT-PCR revealed expression of human Dp 71 but not of Dp 116 or Dp 40. Remarkably, differences were observed in processing of the 3' region of the endogenous mouse and the human transcripts, due to different splicing of exons 71 (absent in human and present in mouse transcript) and 78 (present in human and absent in mouse transcript). The splicing pattern of the human transcript is the same as that of the major Dp 71 (apo-dystrophin-1) product in human blood. The observed splicing differences may be caused by either species-specific exon use and/or by cis-acting factors, e.g. the upstream transcript composition, because we have no evidence for endogenous Dp 71 expression.
Heikoop
J C
JC
MGC-Department of Human Genetics, Leiden University, The Netherlands.
Hogervorst
F B
FB
Meershoek
E J
EJ
Grootscholten
P M
PM
den Dunnen
J T
JT
van Ommen
G J
GJ
D000818:Animals; D001483:Base Sequence; D018244:Chromosomes, Artificial, Yeast; D017931:DNA Primers / Q000235:genetics; D018076:DNA, Complementary / Q000235:genetics; D016189:Dystrophin / Q000031:analogs & derivatives* / Q000235:genetics; D005091:Exons; D005786:Gene Expression Regulation; D018014:Gene Transfer Techniques; D006801:Humans; D006822:Hybrid Cells; D017404:In Situ Hybridization, Fluorescence; D007739:L Cells; D051379:Mice; D008969:Molecular Sequence Data; D009136:Muscular Dystrophies / Q000235:genetics; D016133:Polymerase Chain Reaction; D011401:Promoter Regions, Genetic; D012313:RNA / Q000235:genetics; D012326:RNA Splicing
D017931:DNA Primers; D018076:DNA, Complementary; D016189:Dystrophin; C085339:apo-dystrophin 1; D012313:RNA