Generation and characterization of the <i>jhamt</i> mutants.

<p>(A) Scheme for <i>jhamt</i> targeting. <i>jhamt</i> (black box) is replaced with <i>white</i> (gray box) by homologous recombination of the flanking sequences (white boxes). Red bars represent the primer pairs <i>jhamt-1/jhamt-2</i>; green bars, <i>jhamt-3/jhamt-4</i>; purple bars, <i>jhamt-1/jhamt-5</i>; blue bars, <i>jhamt-6/jhamt-4</i>. (B) Genomic DNA PCR to detect <i>white</i> and <i>jhamt</i> DNA using the above-mentioned primer pairs and genomic DNA extracted from <i>pw25-jhamt</i> and <i>jhamt</i>^<i>1</i>, <i>jhamt</i>^<i>2</i>, and <i>jhamt</i>^<i>3</i> lines. (C) Reverse transcription PCR to detect <i>jhamt</i> mRNA (<i>rp49</i> as the internal control) from <i>pw25-jhamt</i> and <i>jhamt</i>^<i>1</i>, <i>jhamt</i>^<i>2</i>, and <i>jhamt</i>^<i>3</i>. All of the mRNAs were isolated from the brain-RG complexes at 3 hours after initiation of wandering (3hAIW). (D and D’) Immunohistochemistry to detect JHAMT in the CA of <i>w</i>^<i>1118</i> (D) and <i>jhamt</i>^<i>2</i> (D’) at 3hAIW. The red arrow points to the CA showing JHAMT expression. (E) The lethality of <i>w</i>^<i>1118</i> and <i>jhamt</i>^<i>2</i> homozygous mutant during embryonic, larval and pupal stages.