Journal:

Article Title: Cloning, Characterization, and Transcription of Three Laccase Genes from Gaeumannomyces graminis var. tritici , the Take-All Fungus

doi: 10.1128/AEM.68.3.1305-1311.2002

Figure Lengend Snippet: (A) Transcription patterns of the LAC1 gene. RT-PCR was performed with LAC1-specific primers, and total RNA was isolated from G. graminis grown in starvation medium (lane 2), minimal medium supplemented with 400 μm CuSO4 (lane 3), minimal medium without copper (lane 4), PDB (lane 5), LB medium supplemented with 400 μM CuSO4 (lane 6), LB medium without copper (lane 7), and LB medium supplemented with sterile plant homogenate (lane 8). Lane 9 shows RT-PCR with LAC1-specific primers and RNA purified from wheat plants infected with G. graminis; lane 10 shows RT-PCR with LAC1-specific primers and RNA purified from uninfected control wheat plants. Lane 11 shows amplified genomic LAC1, which contains two introns. Lane 1 shows a 100-bp ladder (Promega) as molecular size markers. (B) Transcription patterns of the LAC2 gene. RT-PCR was performed with LAC2-specific primers and total RNA isolated from G. graminis grown in starvation medium (lane 2); minimal medium supplemented with 400 μM CuSO4 (lane 3); minimal medium without copper (lane 5); PDB (lane 7); LB medium supplemented with 400 μm CuSO4 (lane 8); and LB medium without copper (lane 10). Lane 12 shows RT-PCR with primers specific for LAC2 and RNA purified from the wheat plants infected with G. graminis; lane 14 shows RT-PCR with primers specific for LAC2 and RNA purified from the uninfected control wheat plants; and lane 15 shows amplified genomic LAC2, which has no introns. Control reactions without reverse transcriptase are shown in lanes 4, 6, 9, 11, and 13. Lane 1 shows 100-bp ladder molecular size markers. (C) Transcription patterns of the LAC3 gene. RT-PCR was performed with LAC3-specific primers, and total RNA was isolated from wheat plants infected with G. graminis var. tritici (lane 2); G. graminis var. tritici grown in minimal medium supplemented with 30% (vol/vol) sterile plant homogenate (lane 3); minimal medium with 400 μM CuSO4 (lane 6); minimal medium without copper (lane 7); starvation medium (lane 8); PDB (lane 9); LB with 400 μM CuSO4 (lane 10); and LB without copper (lane 11). Lane 4 shows amplified genomic LAC3, which contains an intron. Lane 5 shows RT-PCR with primers specific for LAC3 and RNA purified from the uninfected control wheat plants. Lane 1 shows 100-bp ladder molecular size markers (Promega). (D) Transcription patterns of the POLG gene. RT-PCR was performed with POLG-specific primers and total RNA isolated from G. graminis grown in starvation medium (lane 2); PDB (lane 4); minimal medium supplemented with 400 μM CuSO4 (lane 6); minimal medium without copper (lane 8); LB medium supplemented with 400 μM CuSO4 (lane 10); and LB medium without copper (lane 12). Lane 14 shows RT-PCR with primers specific for POLG and RNA purified from the wheat plants infected with G. graminis; lane 16 shows RT-PCR with primers specific for POLG and RNA purified from the uninfected control wheat plants; lane 17 shows amplified genomic POLG, which has no introns. Control reactions without reverse transcriptase are shown in lanes 3, 5, 7, 9, 11, 13, and 15. Lane 1 shows 100-bp ladder molecular size markers.

Article Snippet: Single-tube Access reverse transcription (RT)-PCR (Promega) was used to monitor the differential transcription of laccase genes.

Techniques: Reverse Transcription Polymerase Chain Reaction, Isolation, Purification, Infection, Amplification