1887

Abstract

For studies of differential gene expression in prokaryotes, methods for synthesizing representative cDNA populations are required. Here, a technique is described for the synthesis of cDNA from the potato pathogens subsp. () and subsp. () using a combination of short oligonucleotide (11-mer) primers that were known to anneal to conserved sequences in the 3′ regions of enterobacterial genes. Specific PCR amplifications with primers designed to anneal to 14 known genes from either or revealed the presence of the corresponding transcripts in cDNA, suggesting that the cDNA represented a broad genomic coverage. cDNA-amplified fragment length polymorphism (cDNA-AFLP) was used to identify differentially expressed genes in , including one that shows significant similarity, at the protein level, to an avirulence gene from pv. . Northern analysis was used to confirm that differentially amplified cDNA fragments were derived from differentially expressed genes. This is the first report of the use of cDNA-AFLP to study differential gene expression in prokaryotes.

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2000-01-01
2019-12-14
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