Use of second-site homologous recombination to demonstrate that Epstein-Barr virus nuclear protein 3B is not important for lymphocyte infection or growth transformation in vitro - PubMed (original) (raw)

Use of second-site homologous recombination to demonstrate that Epstein-Barr virus nuclear protein 3B is not important for lymphocyte infection or growth transformation in vitro

B Tomkinson et al. J Virol. 1992 May.

Abstract

Recombinant Epstein-Barr viruses with a stop codon inserted into the nuclear protein 3B (EBNA 3B) open reading frame were generated by second-site homologous recombination. These mutant viruses infected and growth transformed primary B lymphocytes, resulting in the establishment of lymphoblastoid cell lines (LCLs). Polymerase chain reaction analysis and Southern hybridizations with infected cell DNA demonstrated the presence of the mutant EBNA 3B and the absence of wild-type EBNA 3B. Immunoblot analysis of the LCLs with affinity-purified EBNA 3B antibodies confirmed the absence of EBNA 3B cross-reactive protein. Virus was reactivated from two of these infected LCLs and serially passaged through primary B lymphocytes. The newly infected cells contained only the mutant recombinant virus. No difference was noted between mutant and wild-type recombinants, derived in parallel, in latent (other than EBNA 3B) or lytic cycle-infected cell virus protein expression or in the growth of the latently infected transformed cell lines. These data indicate that the EBNA 3B protein is not critical for primary B-lymphocyte infection, growth transformation, or lytic virus infection in vitro.

PubMed Disclaimer

Similar articles

Cited by

References

    1. Exp Cell Res. 1975 May;92(2):478-84 - PubMed
    1. Nucleic Acids Res. 1981 Oct 24;9(20):5233-52 - PubMed
    1. J Virol. 1992 Feb;66(2):780-9 - PubMed
    1. J Virol. 1991 Dec;65(12):6826-37 - PubMed
    1. J Exp Med. 1990 Jan 1;171(1):345-9 - PubMed

Publication types

MeSH terms

Substances

LinkOut - more resources