Representative segments of display gels of cDNA fragments: left, neutrophils and monocytes exposed to
E coli for 2 hours; right, neutrophils exposed to various bacteria for 2 hours. After incubating the leucocytes with or without bacteria at 37°C for 2 hours, total RNA was extracted, and double-stranded oligo-dT primed cDNA was synthesized by standard methods. The cDNA was digested with a restriction enzyme and ligated with a Y-shaped adapter. Ligated cDNA was amplified by PCR with a phosphorous 32 (32P)–labeled primer complementary to one arm of the Y-shaped adapter and a second primer complementary to the oligo-dT primer and containing one of the 12 possible dinucleotide extensions on its 3′-end, as previously described.1 2 Ec indicates E coli K12; Yp (KIM5), Y pestis KIM5 (pCD1+); Yp (KIM6), Y pestis KIM6 (pCD1−); N, neutrophil; and M, monocyte. Gene symbols are the same as those described in Tables 4-6, except for the following:PAI2, plasminogen activator inhibitor, type II (arginine-serpin) (GenBank Accession no. Y00630); RPL3, ribosomal protein L3 (X73460); and RPS4X, ribosomal protein S4, X-linked (M58458).
