Summary
A total of 50 different ACTBP2 (human betaactin related pseudogene) alleles and the cell line K562 were sequenced and analysed. Sequence data revealed not only length polymorphism but also a highly variable sequence polymorphism. 2 different sequence structures were characterised. Type I (223–259bp) contained the known regular 4bp repeat AAAG. Type II (265–309 bp) revealed a further hexanucleotide unit AAAAAG in addition to the common AAAG which only occurs once in the repeat region. The position of this insertion showed considerable variation. To obtain a regular spaced allelic ladder 20 of the sequenced alleles were selected. Denaturing gels and high resolution/non-denaturing gels were compared and striking differences could be seen between the 2 gel systems. Separation of the alleles on a 6% denaturing gel and analysis on the ABI 373A Sequencer revealed fragment sizes which corresponded to the sequencing data but were in general 6–10bp longer. In contrast, in non-denaturing gels some alleles showed different electrophoretic mobilities compared to the sequenced allelic ladder which could indicate different fragment length and/or different sequence structure.
Zusammenfassung
50 verschiedene ACTBP2 Allele und die Zell-Linie K562 wurden sequenziert und analysiert. Die Sequenzierergebnisse zeigen nicht nur einen Längensondern auch einen hoch variablen Sequenzpolymorphismus. 2 verschiedene Sequenzstrukturen wurden charakterisiert. Typ 1 (223–259bp) enthält den regulären 4bp-Repeat AAAG. Im Typ II (265–309 bp) tritt zusätzlich die Hexanukleotideinheit AAAAAG auf, deren Position innerhalb der Repeatregion stark variabel ist. 20 sequenzierte Allele wurden für die Konstruktion einer allelischen Leiter ausgewählt. Denaturierende und hochauflösende, nicht denaturierende Gele wurden ausgewertet und Unterschiede im Wanderungsverhalten der Allele festgestellt. In denaturierenden Gelen ist die elektrische Mobilität abhängig von den Fragmentlängen der Allele. Dagegen zeigen in nicht denaturierenden Gelen einige Allele unterschiedliches Wanderungsverhalten im Vergleich zur sequenzierten allelischen Leiter, welches durch unterschiedliche Fragmentlängen und/oder unterschiedliche Sequenzstrukturen erklärt werden kann.
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Möller, A., Brinkmann, B. Locus ACTBP2 (SE33). Int J Leg Med 106, 262–267 (1994). https://doi.org/10.1007/BF01225417
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DOI: https://doi.org/10.1007/BF01225417