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Journal of Clinical Microbiology, December 2000, p. 4310-4314, Vol. 38, No. 12
0095-1137/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Application of Representational Difference Analysis to Genomic Fragments of Marek's Disease Virus

Daiji Endoh,1,* Kyoung-Oh Cho,2,dagger Kenji Tsukamoto,3 Toshifumi Morimura,2 Yasuhiro Kon,4 and Masanobu Hayashi1

Department of Veterinary Radiology, Rakuno Gakuen University, Ebetsu,1 Department of Disease Control,2 and Laboratory of Experimental Animal Science,4 Graduate School of Veterinary Medicine, Hokkaido University, Hokkaido, and Laboratory of Viral Pathogenesis, National Institute of Animal Health, Tsukuba,3 Japan

Received 2 May 2000/Returned for modification 16 August 2000/Accepted 21 September 2000

A rapid and simple method for isolation of DNA fragments of Marek's disease virus (MDV) based on representational difference analysis (RDA) was developed. Multiple viral DNA fragments, the sizes of which were restricted to 0.3 to 3.5 kbp, were simultaneously amplified after subtraction of chicken DNA from BamHI-, BglII-, EcoRI-, HindIII-, or XhoI-digested DNA fragments of MDV-infected cells. Nucleotide sequence of two RDA-derived fragments coincided with the sequence determined from direct sequencing of the MDV genome. We detected an interstrain difference in the size of restriction enzyme-digested fragments on agarose gel. This method was used on a single feather pulp to generate sufficient MDV DNA for cloning.

* Corresponding author. Mailing address: Department of Veterinary Radiology, Rakuno Gakuen University, Ebetsu 069-8501, Japan. Phone: 81-11-388-4847. Fax: 81-11-387-5890. E-mail: dendoh{at}rakuno.ac.jp.

dagger Present address: Department of Veterinary Pathology, College of Veterinary Medicine, Chonnam National University, Kwangju, South Korea.

Journal of Clinical Microbiology, December 2000, p. 4310-4314, Vol. 38, No. 12
0095-1137/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.




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