Sequence analysis of the <i>MYD88</i> gene in Waldenström’s macroglobulinemia.

<div><p>(A) Sequencing revealed a T to C transition resulting in a leucine to proline substitution at amino acid position 265 (WM5).</p> <p>(B) Wild-type sequences (T) are shown as a control (WM2). </p> <p>(C) Direct sequencing showed both wild-type and mutant alleles in WM5 and WM9, and the wild-type allele only in WM2. </p> <p>(D) Sensitivity of direct sequencing. L265P-positive DNA (WM5) was diluted into wild-type DNA (WM3) before amplification. Aberrant bands were detected in samples containing 10% or more of the L265P mutation.</p></div

Allele-specific polymerase chain reaction (AS-PCR) of the <i>MYD88</i> gene in Waldenström’s macroglobulinemia and lymphoma.

<div><p>Ten μl of PCR products was separated by electrophoresis through a 2% agarose gel, stained with ethidium bromide, and visualized by ultraviolet illumination. The size of the products is indicated on the left.</p> <p>(A) Sensitivity of AS-PCR. L265P-positive DNA (WM5) was diluted into wild-type DNA (WM2) before amplification. Mutations were detected in samples containing 0.1% or more of the L265P mutation. Lane 1, 100 bp ladder; lane 2, 0%; lane 3, 0.1%; lane 4, 0.5%; lane 5, 1%; lane 6, 0%; lane 7, 0.1%; lane 8, 0.5%; lane 9, 1%.</p> <p>(B) Aberrant bands were detected in 12 of 16 samples from WM patients (lanes 2, 5, 6, 8, 9, 10, 11, 12, 14, 15, 16, and 17). Lane 1, 100 bp ladder; lane 2, WM1; lane 3, WM2; lane 4, WM3; lane 5, WM4; lane 6, WM5; lane 7, WM6; lane 8, WM7; lane 9, WM8; lane 10, WM9; lane 11, WM10; lane 12, WM11; lane 13, WM12, lane 14, WM13; lane 15, WM14; lane 16, WM15; lane 17, WM16.</p> <p>(C) Aberrant bands were detected in 6 of 10 samples from WM patients (lanes 2, 3, 6, 7, 8, and 10) and 2 of 3 non-Hodgkin’s lymphoma patients (lanes 12 and 13). Lane 1, 100 bp ladder; lane 2, WM17; lane 3, WM18; lane 4, WM19; lane 5, WM20; lane 6, WM21; lane 7, WM22; lane 8, WM23; lane 9, WM24; lane 10, WM25; lane 11, NHL1; lane 12, NHL2; lane 13, NHL3; lane 14, normal lymphocyte; lane 15, water.</p></div

Location of the analyzed mutation and a schema of the involved pathway.

<p>TIRAP, TIR domain containing adaptor protein; IRAK, IL-1 receptor-associated kinase; BTK, Bruton’s tyrosine kinase; TRAF, tumor necrosis factor receptor-associated factor; TAK, TGF-β-activated kinase; TAB, TAK binding protein; IκB, inhibitor κB; IKK, IκB kinase; NF-κB, nuclear factor-κB.</p