Intracellular expression and processing of foot-and-mouth disease virus capsid precursors using vaccinia virus vectors: Influence of the L protease
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Intracellular expression and processing of foot-and-mouth disease virus capsid precursors using vaccinia virus vectors : Influence of the L protease. / Belsham, Graham J.; Brangwyn, Julia K.; Ryan, Martin D.; Abrams, Charles C.; King, Andrew M.Q.
In: Virology, Vol. 176, No. 2, 06.1990, p. 524-530.Research output: Contribution to journal › Journal article › Research › peer-review
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TY - JOUR
T1 - Intracellular expression and processing of foot-and-mouth disease virus capsid precursors using vaccinia virus vectors
T2 - Influence of the L protease
AU - Belsham, Graham J.
AU - Brangwyn, Julia K.
AU - Ryan, Martin D.
AU - Abrams, Charles C.
AU - King, Andrew M.Q.
N1 - Funding Information: This work was supported in part by the European Community Biotechnology Action Programme Research Contract BAP 01 1g -UK(H) and in part by Pfizer. We thank B. Moss and G. Smith for supplying us with the vTF7-3 vaccinia recombinant and plasmids pGS20 and pAR2529. We thank John McCauley for synthesizing the oligonucleotides and Chris Bostock for critical reading of the manuscript. We also thank Doros Panayi for photography and Fiona Berrie for typing.
PY - 1990/6
Y1 - 1990/6
N2 - cDNA cassettes of FMDV have been constructed which encode the capsid precursor (P1-2A) alone or with the proteases L and 3C which are required for processing of this precursor to the products 1 AB, 1 C, and 1 D. These cassettes have been analyzed using in vitro transcription and translation reactions and within cells using recombinant vaccinia viruses. Processing of the precursors occurred more efficiently in cells than in cell-free systems but similar properties were observed. It was not possible to isolate recombinant vaccinia viruses containing FMDV cassettes which included the intact coding sequence for the L protein. Deletion of part of the L sequence, which abolished its proteolytic activity, also abolished this incompatibility with vaccinia virus. The vaccinia recombinant, vTF7-3, which expresses the bacteriophage T7 RNA polymerase was used in transient expression studies using plasmids containing a T7 promoter upstream of the FMDV cassettes. Under these conditions it was possible to coexpress L, P1-2A, and 3C in the vaccinia-infected cells; each of the proteolytic activities was observed and correctly processed 1AB, 1C, and 1D were produced.
AB - cDNA cassettes of FMDV have been constructed which encode the capsid precursor (P1-2A) alone or with the proteases L and 3C which are required for processing of this precursor to the products 1 AB, 1 C, and 1 D. These cassettes have been analyzed using in vitro transcription and translation reactions and within cells using recombinant vaccinia viruses. Processing of the precursors occurred more efficiently in cells than in cell-free systems but similar properties were observed. It was not possible to isolate recombinant vaccinia viruses containing FMDV cassettes which included the intact coding sequence for the L protein. Deletion of part of the L sequence, which abolished its proteolytic activity, also abolished this incompatibility with vaccinia virus. The vaccinia recombinant, vTF7-3, which expresses the bacteriophage T7 RNA polymerase was used in transient expression studies using plasmids containing a T7 promoter upstream of the FMDV cassettes. Under these conditions it was possible to coexpress L, P1-2A, and 3C in the vaccinia-infected cells; each of the proteolytic activities was observed and correctly processed 1AB, 1C, and 1D were produced.
UR - http://www.scopus.com/inward/record.url?scp=0025307235&partnerID=8YFLogxK
U2 - 10.1016/0042-6822(90)90022-J
DO - 10.1016/0042-6822(90)90022-J
M3 - Journal article
C2 - 2161149
AN - SCOPUS:0025307235
VL - 176
SP - 524
EP - 530
JO - Virology
JF - Virology
SN - 0042-6822
IS - 2
ER -
ID: 381223513