Infectious foot-and-mouth disease virus derived from a cloned full-length cDNA

Abstract

A full-length cDNA plasmid of foot-and-mouth disease virus has been constructed. RNA synthesized in vitro by means of a bacteriophage SP6 promoter inserted in front of the cDNA led to the production of infectious particles upon transfection of BHK-21 ceUs. These particles were also found to be highly infectious for primary bovine kidney cells as well as for baby mice. The difficulty in cloning the foot-and-mouth disease virus cytidyl tract in Escherichia coli was circumvented by joining two separate cloned parts, representing the S and L fragments of the genome, and, in a second step, inserting a dC-dG homopolymer. Homopolymeric sequences of up to 25 cytidyl residues did not lead to the production of virus. Replicons containing poly(C) tracts long enough to permit virus replication were first established in yeast cells. One of these constructs could also be maintained in E. coli and was used to produce infectious RNA in vitro. The length of the poly(C) sequence in this cDNA plasmid was 32 nucleotides. However, the poly(C) tracts of two recombinant viruses found in transfected BHK-21 cells were 60 and 80 nucleotides long, respectively. Possible mechanisms leading to the enlargement of the poly(C) tract during virus replication are discussed.

The Microbial Rosetta Stone Database: A compilation of global and emerging infectious microorganisms and bioterrorist threat agents

“…Chemical synthesis of RNA viruses would eliminate the need for the bioterrorist to obtain stock cultures and would allow creation of any wild-type or mutant strain of virtually any virus. Other positive-strand RNA viruses ( Figure 6) that have been reconstituted from cloned DNA samples include Kunjin virus [59], human rhinovirus [60], foot-and-mouth disease virus [61], transmissible gastroenteritis virus, a coronavirus [62,63], all three genera of the family Flaviviridae [64], and a variety of plant and insect viruses [65,66]. Thus, the positivestranded viruses must be considered a high-risk area for bioengineering, where the relative risk for each virus must be considered roughly equivalent to the terrorist potential inherent to the virus.…”

Distinctive features of foot-and-mouth disease virus, a member of the picornavirus family; aspects of virus protein synthesis, protein processing and structure

“…The presence of the poly C tract within FMDV inhibited progress towards the production of infectious cDNA for FMDV. However an infectious cDNA copy of FMDV has now been constructed using the O 1K strain (Zibert et al, 1990); this infectious clone contains just 32 C residues in the poly C tract. This number was something of a compromise between the number that could be stably maintained within a bacterial plasmid and that apparently required for viability.…”

“…1. The complete nucleotide (nt) sequence (8400 nt) of the O1Kaufbeuren (O1K) strain of FMDV has been determined (Forss et al, 1984;Zibert et al, 1990). The 5' non-coding region (NCR) of FMDV is exceptionally long, it contains about 1300 nt; this may be compared with PV containing about 740 nt and typical cellular mRNAs which have 5'NCRs of 50-I00 nt.…”

Pathogenesis of Theiler's Murine Encephalomyelitis Virus

“…In addition to sequence comparisons to determine important areas within the genome, studies using infectious complementary DNA clones have become very powerful. Infectious transcripts for different picornaviruses have been prepared: polioviruses (Racaniello and Baltimore, 1981;Omata et al, 1984;Kaplan et al, 1985), human rhinovirus type 14 (Mizutani and Colonno, 19851, coxsackie B3 virus (Kandolf and Hofschneider, 1985), EMCV (Naviaux et al, 19901, hepa-titis A (Cohen et al, 1987), and foot-and-mouth disease virus (Zibert et al, 1990). Infectious transcripts for members of both TMEV subgroups DA (Roos et al, 1989131, BeAn (Calenoff et al, 1990), and the highly virulent GDVII (Tangy et al, 1989) have been constructed.…”