Thc immediate early genc 1 (IE1) is thc first gene tobe cxpressed following thc cntry ofthc human cytomcgalovirus (HCMV) into the cell and it docs not requirc prior protcin synthesis for its expression. Thereforc, the IE1 gcnc is a potential candidatc for the devclopment of probcs to detcct HCMV in various states of infection. Using strand-specific 32P- or digoxigenin-labeled riboprobes derivcd from an cxon-specific subgenomic fragment of thc HCMV Towne IE1 genc, wc perforrncd Northern blot analysis and RNA in situ hybridization on HCMV-infected human (permissive cells) and mouse (nonpcrrnissivc cclls) fibroblasts and on 10 formalin-fixed paraffin-cmbcddcd scctions ofhuman tissue. By Northem blot analysis and by in situ hybridization, expression of thc 2.0-kb IE1 gcne was found in permissive as wcll as in nonpermissive infcctions. Specific nuclear and cytoplasmic hybridization was found at 5, 10, 24, and 72 h alter infection in human fibroblasts. In comparison, hybridization was first dctectcd at 10 h aftcr infcction in mouse fibroblasts. Hybridization with thc JE 1 probe was detected in cclls with and without cytopathic changcs in the formalin-fixed paraffin-cmbcddcd HCMV-infected human tissues. Hybridization patterns of thc IE1 riboprobe were compared to thosc of the HCMV 2.7-kb major early β-riboprobc which we havc prcviously described [Am J Pathol 141: 1247-1254; 1992]. Although both riboprobcs hybridize to their rcspcctive targct scquenccs in the consecutivc tissue sections, the pattcrns of hybridization arc different. On occasion, sections ofHCMV-infected human tissue showing no specific hybridization for the 2.7-kb riboprobc will show spccific in situ hybridization when using thc IE! riboprobc. Our rcsults suggcst that RNA in situ hybridization with a probe dirccted at thc IE! transcripts is an effcctivc method ofdctecting early and late stages ofboth permissive and nonpermissivc HCMV infcctions.

Keywords:
Cytomegalovirus, Immediate early genc 1, Major early β-gene, 2.7 kb, Hybridization in situ, Permissive infection, Nonpermissive infection
This content is only available via PDF.
1997
Copyright / Drug Dosage / Disclaimer
Copyright: All rights reserved. No part of this publication may be translated into other languages, reproduced or utilized in any form or by any means, electronic or mechanical, including photocopying, recording, microcopying, or by any information storage and retrieval system, without permission in writing from the publisher.
Drug Dosage: The authors and the publisher have exerted every effort to ensure that drug selection and dosage set forth in this text are in accord with current recommendations and practice at the time of publication. However, in view of ongoing research, changes in government regulations, and the constant flow of information relating to drug therapy and drug reactions, the reader is urged to check the package insert for each drug for any changes in indications and dosage and for added warnings and precautions. This is particularly important when the recommended agent is a new and/or infrequently employed drug.
Disclaimer: The statements, opinions and data contained in this publication are solely those of the individual authors and contributors and not of the publishers and the editor(s). The appearance of advertisements or/and product references in the publication is not a warranty, endorsement, or approval of the products or services advertised or of their effectiveness, quality or safety. The publisher and the editor(s) disclaim responsibility for any injury to persons or property resulting from any ideas, methods, instructions or products referred to in the content or advertisements.
You do not currently have access to this content.