Figure 3 | Oncogene

Figure 3

From: AF15q14, a novel partner gene fused to the MLL gene in an acute myeloid leukaemia with a t(11;15)(q23;q14)

Figure 3

Schematic representation of the AF15q14 locus and the repeat motif encoded by exon 10 of AF15q14. (a) Human AF15q14 cDNA was analysed by a combination of rapid amplification of 5′/3′cDNA ends from HeLa cells poly(A)+ RNA (5′/3′ RACE kit, Roche), from testis cDNA (Marathon-Ready cDNA, Clontech) and by the screening of a human HeLa cDNA library (Clontech) with the AF15c probe. A control with a tube containing RNA, but without reverse transcriptase, was performed in order to rule out possible genomic amplification. PCR products were subcloned into the TA cloning vector (InVitrogen). Genomic DNA fragments from normal chromosome 15 were obtained either by placenta-library screening and analysis of the phages subcloned into Bluescript SK(−) vector (Stratagene) or by direct sequencing of PCR-amplified genomic DNA fragments (Expand long Template PCR System, Roche). Sequence analysis of cDNA from both strands and their genomic counterparts was performed by the dideoxynucleotide method. Comparison of the human cDNA and genomic DNA AF15q14 sequences showed that the AF15q14 gene extended for more than 35 kb, and contained at least 10 exons. Analysis of overlapping cDNA clones allowed us to sequence 5925 bp of the AF15q14 transcript (GenBank accession No. AF248041). The breakpoint on the AF15q14 gene is indicated by a vertical arrow. The sequence encoding the NLS-BP is shown by a black dot. Ex: exon (unknown 3′end of AF15q14 exon 10 is indicated by dotted line); Int: intron; CEN: centromere; TEL: telomere. (b) Analysis of the predictive AF15q14 protein (1833 amino acids) with the Antheprot program (Geourjon and Deleage, 1995) revealed a repeat motif of about 105 amino acids between residues 859 and 1175. These motifs, though sharing no particular specificity, had about 73% of amino-acids identity

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