PMID- 10222241 OWN - NLM STAT- MEDLINE DCOM- 19990527 LR - 20061115 IS - 0006-291X (Print) IS - 0006-291X (Linking) VI - 258 IP - 1 DP - 1999 Apr 29 TI - Search for genes potentially involved in Mycobacterium tuberculosis virulence by mRNA differential display. PG - 94-101 AB - An mRNA differential display (DD) assay was developed to compare gene expression between Mycobacterium tuberculosis H37Rv and its avirulent mutant H37Ra. The DD protocol made use of an oligo(dT) to prime reverse-transcriptase (RT)-dependent transcription of poly-A tailed mRNAs and a PCR amplification of the RT products by using ten 12-base arbitrary primers in all their pair combinations. This analysis yielded 745 and 708 bands, including 52 and 15 differentially generated bands, in the strains H37Rv and H37Ra, respectively. Six cDNAs that appeared to be expressed in H37Rv, but not in H37Ra, were reamplified and cloned and at least 10 inserts were sequenced for each cloned cDNA. After resolving discrepant results, 6 inserts were found highly homologous to M. tuberculosis H37Rv genes. Three of these, i.e., genes Rv2770c, Rv1345, and Rv0288, coding respectively for a member of the PPE protein family, a probable polyketide synthase, and a member of the protein family containing ESAT-6, have been predictively associated to immunological or pathogenetic aspects of M. tuberculosis infection; the other genes, i.e., Rv2336, Rv1320c, and Rv2819c, code for proteins with unknown functions. These results show that mRNA DD methodology can represent a potential tool for investigation of M. tuberculosis gene expression. CI - Copyright 1999 Academic Press. FAU - Rindi, L AU - Rindi L AD - Biotecnologie Mediche, Infettivologia ed Epidemiologia, Universita di Pisa, Pisa, I-56127, Italy. FAU - Lari, N AU - Lari N FAU - Garzelli, C AU - Garzelli C LA - eng PT - Journal Article PT - Research Support, Non-U.S. Gov't PL - United States TA - Biochem Biophys Res Commun JT - Biochemical and biophysical research communications JID - 0372516 RN - 0 (DNA Primers) RN - 0 (RNA, Messenger) SB - IM MH - Base Sequence MH - Cloning, Molecular MH - DNA Primers MH - *Genes, Bacterial MH - Mycobacterium tuberculosis/genetics/*pathogenicity MH - RNA, Messenger/*genetics MH - Virulence/*genetics EDAT- 1999/05/01 00:00 MHDA- 1999/05/01 00:01 CRDT- 1999/05/01 00:00 PHST- 1999/05/01 00:00 [pubmed] PHST- 1999/05/01 00:01 [medline] PHST- 1999/05/01 00:00 [entrez] AID - S0006-291X(99)90591-0 [pii] AID - 10.1006/bbrc.1999.0591 [doi] PST - ppublish SO - Biochem Biophys Res Commun. 1999 Apr 29;258(1):94-101. doi: 10.1006/bbrc.1999.0591.