An amelogenin minigene to study alternative splicing.
Diversity in gene expression is commonly observed as a result of alternative splicing of RNA transcripts. This is true in the case of amelogenin, one of the enamel matrix proteins. Our hypothesis is that additional amelogenin mRNA transcripts are generated in vivo, but these transcripts have yet to be observed because of the limitations of currently used detection methodologies. For this study our objective was to create an amelogenin minigene to study amelogenin RNA splicing events in cell lines of diverse character. Mouse genomic DNA was used as a PCR template to amplify the amelogenin DNA sequence spanning exons 2-7. The resulting PCR-generated DNA was subcloned in an expression vector. This resulting amelogenin minigene was shown to be functionally active by transfection into multiple cell lines. We have successfully cloned an amelogenin minigene, and as a result we describe and discuss novel amelogenin alternatively spliced transcripts.
Shapiro
Jason L
JL
Center for Craniofacial Molecular Biology, University of Southern California School of Dentistry, Los Angeles, 90033, USA.
Wang
Hongjun
H
Wen
Xin
X
Tannukit
Sissada
S
Paine
Michael L
ML
D017398:Alternative Splicing; D053523:Amelogenin; D000595:Amino Acid Sequence; D000818:Animals; D002460:Cell Line; D003001:Cloning, Molecular; D003746:Dental Enamel Proteins / Q000235:genetics*; D005091:Exons; D051379:Mice; D008969:Molecular Sequence Data; D012322:RNA Precursors / Q000235:genetics; D016415:Sequence Alignment
D053523:Amelogenin; C507200:Amelx protein, mouse; D003746:Dental Enamel Proteins; D012322:RNA Precursors