Nucleotide sequence of the capsid protein cistrons from six potato virus Y (PVY) isolates infecting tobacco

Sudarsono, S. L. Woloshuk, Zhongguo Xiong, G. M. Hellmann, E. A. Wernsman, A. K. Weissinger, S. A. Lommel

Research output: Contribution to journalArticle

21 Citations (Scopus)

Abstract

Complementary DNA libraries representing the capsid protein cistron of the potato virus Y (PVY) isolate 'Chilean', 'Hungarian', MsNr, NsNr, O, and 'Potato US' were synthesized and used as template for polymerase chain reaction (PCR) amplification. An AUG codon for initiating a discrete capsid protein (CP) open reading frame was embedded upstream of the first codon of the CP cistrons. PCR-amplified products of the expected size of 0.8 kilo bases were cloned into the transcription vector pBS(+). The fidelity of each PCR-amplified PVY CP cistron was tested by transcribing recombinant plasmids in vitro and translating the transcripts in two cell free translation systems. Translation analysis of in vitro transcribed PVY CP cistrons consistently yielded a polypeptide co-migrating with authentic CP that was immunoprecipitated by anti PVY 'Chilean' antibodies. The nucleotide sequence of each capsid protein gene was determined by dideoxy sequence analysis. Each capsid protein gene was determined to be 801 nucleotides in length, encoding a deduced protein of 267 amino acids with calculated Mr ranging from 29 799 to 29 980. The nucleic acid sequence similarity between the six isolates ranged between 89 to 97% and the amino acid similarity between 91 to 99%. The high level of amino acid sequence similarity confirms the classification of these viruses as isolates of PVY.

Original languageEnglish (US)
Pages (from-to)161-170
Number of pages10
JournalArchives of Virology
Volume132
Issue number1-2
DOIs
StatePublished - Mar 1993

Fingerprint

Potyvirus
Capsid Proteins
Tobacco
Genes
Codon
Polymerase Chain Reaction
Amino Acids
Cell-Free System
Solanum tuberosum
Gene Library
Nucleic Acids
Open Reading Frames
Sequence Analysis
Amino Acid Sequence
Plasmids
Nucleotides
Complementary DNA
Viruses
Peptides
Antibodies

ASJC Scopus subject areas

  • Applied Microbiology and Biotechnology
  • Genetics

Cite this

Sudarsono, Woloshuk, S. L., Xiong, Z., Hellmann, G. M., Wernsman, E. A., Weissinger, A. K., & Lommel, S. A. (1993). Nucleotide sequence of the capsid protein cistrons from six potato virus Y (PVY) isolates infecting tobacco. Archives of Virology, 132(1-2), 161-170. https://doi.org/10.1007/BF01309850

Nucleotide sequence of the capsid protein cistrons from six potato virus Y (PVY) isolates infecting tobacco. / Sudarsono; Woloshuk, S. L.; Xiong, Zhongguo; Hellmann, G. M.; Wernsman, E. A.; Weissinger, A. K.; Lommel, S. A.

In: Archives of Virology, Vol. 132, No. 1-2, 03.1993, p. 161-170.

Research output: Contribution to journalArticle

Sudarsono, Woloshuk, SL, Xiong, Z, Hellmann, GM, Wernsman, EA, Weissinger, AK & Lommel, SA 1993, 'Nucleotide sequence of the capsid protein cistrons from six potato virus Y (PVY) isolates infecting tobacco', Archives of Virology, vol. 132, no. 1-2, pp. 161-170. https://doi.org/10.1007/BF01309850
Sudarsono ; Woloshuk, S. L. ; Xiong, Zhongguo ; Hellmann, G. M. ; Wernsman, E. A. ; Weissinger, A. K. ; Lommel, S. A. / Nucleotide sequence of the capsid protein cistrons from six potato virus Y (PVY) isolates infecting tobacco. In: Archives of Virology. 1993 ; Vol. 132, No. 1-2. pp. 161-170.
@article{a8095a057d594d31a92426d268efccfe,
title = "Nucleotide sequence of the capsid protein cistrons from six potato virus Y (PVY) isolates infecting tobacco",
abstract = "Complementary DNA libraries representing the capsid protein cistron of the potato virus Y (PVY) isolate 'Chilean', 'Hungarian', MsNr, NsNr, O, and 'Potato US' were synthesized and used as template for polymerase chain reaction (PCR) amplification. An AUG codon for initiating a discrete capsid protein (CP) open reading frame was embedded upstream of the first codon of the CP cistrons. PCR-amplified products of the expected size of 0.8 kilo bases were cloned into the transcription vector pBS(+). The fidelity of each PCR-amplified PVY CP cistron was tested by transcribing recombinant plasmids in vitro and translating the transcripts in two cell free translation systems. Translation analysis of in vitro transcribed PVY CP cistrons consistently yielded a polypeptide co-migrating with authentic CP that was immunoprecipitated by anti PVY 'Chilean' antibodies. The nucleotide sequence of each capsid protein gene was determined by dideoxy sequence analysis. Each capsid protein gene was determined to be 801 nucleotides in length, encoding a deduced protein of 267 amino acids with calculated Mr ranging from 29 799 to 29 980. The nucleic acid sequence similarity between the six isolates ranged between 89 to 97{\%} and the amino acid similarity between 91 to 99{\%}. The high level of amino acid sequence similarity confirms the classification of these viruses as isolates of PVY.",
author = "Sudarsono and Woloshuk, {S. L.} and Zhongguo Xiong and Hellmann, {G. M.} and Wernsman, {E. A.} and Weissinger, {A. K.} and Lommel, {S. A.}",
year = "1993",
month = "3",
doi = "10.1007/BF01309850",
language = "English (US)",
volume = "132",
pages = "161--170",
journal = "Archives of Virology",
issn = "0304-8608",
publisher = "Springer Wien",
number = "1-2",

}

TY - JOUR

T1 - Nucleotide sequence of the capsid protein cistrons from six potato virus Y (PVY) isolates infecting tobacco

AU - Sudarsono,

AU - Woloshuk, S. L.

AU - Xiong, Zhongguo

AU - Hellmann, G. M.

AU - Wernsman, E. A.

AU - Weissinger, A. K.

AU - Lommel, S. A.

PY - 1993/3

Y1 - 1993/3

N2 - Complementary DNA libraries representing the capsid protein cistron of the potato virus Y (PVY) isolate 'Chilean', 'Hungarian', MsNr, NsNr, O, and 'Potato US' were synthesized and used as template for polymerase chain reaction (PCR) amplification. An AUG codon for initiating a discrete capsid protein (CP) open reading frame was embedded upstream of the first codon of the CP cistrons. PCR-amplified products of the expected size of 0.8 kilo bases were cloned into the transcription vector pBS(+). The fidelity of each PCR-amplified PVY CP cistron was tested by transcribing recombinant plasmids in vitro and translating the transcripts in two cell free translation systems. Translation analysis of in vitro transcribed PVY CP cistrons consistently yielded a polypeptide co-migrating with authentic CP that was immunoprecipitated by anti PVY 'Chilean' antibodies. The nucleotide sequence of each capsid protein gene was determined by dideoxy sequence analysis. Each capsid protein gene was determined to be 801 nucleotides in length, encoding a deduced protein of 267 amino acids with calculated Mr ranging from 29 799 to 29 980. The nucleic acid sequence similarity between the six isolates ranged between 89 to 97% and the amino acid similarity between 91 to 99%. The high level of amino acid sequence similarity confirms the classification of these viruses as isolates of PVY.

AB - Complementary DNA libraries representing the capsid protein cistron of the potato virus Y (PVY) isolate 'Chilean', 'Hungarian', MsNr, NsNr, O, and 'Potato US' were synthesized and used as template for polymerase chain reaction (PCR) amplification. An AUG codon for initiating a discrete capsid protein (CP) open reading frame was embedded upstream of the first codon of the CP cistrons. PCR-amplified products of the expected size of 0.8 kilo bases were cloned into the transcription vector pBS(+). The fidelity of each PCR-amplified PVY CP cistron was tested by transcribing recombinant plasmids in vitro and translating the transcripts in two cell free translation systems. Translation analysis of in vitro transcribed PVY CP cistrons consistently yielded a polypeptide co-migrating with authentic CP that was immunoprecipitated by anti PVY 'Chilean' antibodies. The nucleotide sequence of each capsid protein gene was determined by dideoxy sequence analysis. Each capsid protein gene was determined to be 801 nucleotides in length, encoding a deduced protein of 267 amino acids with calculated Mr ranging from 29 799 to 29 980. The nucleic acid sequence similarity between the six isolates ranged between 89 to 97% and the amino acid similarity between 91 to 99%. The high level of amino acid sequence similarity confirms the classification of these viruses as isolates of PVY.

UR - http://www.scopus.com/inward/record.url?scp=0027350880&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0027350880&partnerID=8YFLogxK

U2 - 10.1007/BF01309850

DO - 10.1007/BF01309850

M3 - Article

C2 - 8352655

AN - SCOPUS:0027350880

VL - 132

SP - 161

EP - 170

JO - Archives of Virology

JF - Archives of Virology

SN - 0304-8608

IS - 1-2

ER -