Cloning and physical mapping of DNA complementary to potato leafroll virus RNA

Smith, O P

Title: Cloning and physical mapping of DNA complementary to potato leafroll virus RNA

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Abstract

Potato leafroll virus (PLRV) was aphid-transmitted from potato (Solanum tuberosum cultivar Russett Burbank) to ground cherry (Physalis floridana), where it was maintained by serial aphid transmission. Serological and plant differential tests indicated that the isolate was not contaminated with beet western yellows virus. Purified PLRV RNA was poly(A)-tailed in vitro and used as a template for reverse transcriptase, primed with oligo(dT). Alkaline gel electrophoresis of /sup 32/P-labeled first-strand complementary DNA (cDNA) indicated a major size range of 0.1 to 3.5 kilobases (kb). A small percentage of transcripts corresponded to full length PLRV RNA. Following RNase H and DNA polymerase I-mediated second strand synthesis, double-stranded cDNA was cloned into the Pst I site of the plasmid pUC9 using oligo (dC)-oligo(dG) tailing methodology. Escherichia coli JM109 transformants were screened with first-strand /sup 32/P-cDNA in colony hybridization experiments to confirm that recombinants contained PLRV-specific sequences.

Authors:: Smith, O P

Publication Date:: Thu Jan 01 00:00:00 EST 1987

Research Org.:: Texas A and M Univ., College Station (USA)

OSTI Identifier:: 7187998

Resource Type:: Thesis/Dissertation

Resource Relation:: Other Information: Thesis (Ph. D.)

Country of Publication:: United States

Language:: English

Subject:: 59 BASIC BIOLOGICAL SCIENCES; DNA; HYBRIDIZATION; VIRUSES; DNA SEQUENCING; DNA-CLONING; DNA POLYMERASES; ELECTROPHORESIS; PHOSPHORUS 32; RECOMBINANT DNA; RNA; RNA-ASE; SOLANUM TUBEROSUM; TRACER TECHNIQUES; TRANSCRIPTION; BETA DECAY RADIOISOTOPES; BETA-MINUS DECAY RADIOISOTOPES; CLONING; DAYS LIVING RADIOISOTOPES; ENZYMES; ESTERASES; HYDROLASES; ISOTOPE APPLICATIONS; ISOTOPES; LIGHT NUCLEI; MICROORGANISMS; NUCLEI; NUCLEIC ACIDS; NUCLEOTIDYLTRANSFERASES; ODD-ODD NUCLEI; ORGANIC COMPOUNDS; PARASITES; PHOSPHODIESTERASES; PHOSPHORUS ISOTOPES; PHOSPHORUS-GROUP TRANSFERASES; PLANTS; POLYMERASES; RADIOISOTOPES; SOLANUM; STRUCTURAL CHEMICAL ANALYSIS; TRANSFERASES; 550201* - Biochemistry- Tracer Techniques

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                    Smith, O P. Cloning and physical mapping of DNA complementary to potato leafroll virus RNA.  United States: N. p., 1987. 
        Web.

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                    Smith, O P. Cloning and physical mapping of DNA complementary to potato leafroll virus RNA.  United States.

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                    Smith, O P. 1987.  
        "Cloning and physical mapping of DNA complementary to potato leafroll virus RNA".  United States.

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@article{osti_7187998,

  title        = {Cloning and physical mapping of DNA complementary to potato leafroll virus RNA},

  author       = {Smith, O P},

  abstractNote = {Potato leafroll virus (PLRV) was aphid-transmitted from potato (Solanum tuberosum cultivar Russett Burbank) to ground cherry (Physalis floridana), where it was maintained by serial aphid transmission. Serological and plant differential tests indicated that the isolate was not contaminated with beet western yellows virus. Purified PLRV RNA was poly(A)-tailed in vitro and used as a template for reverse transcriptase, primed with oligo(dT). Alkaline gel electrophoresis of /sup 32/P-labeled first-strand complementary DNA (cDNA) indicated a major size range of 0.1 to 3.5 kilobases (kb). A small percentage of transcripts corresponded to full length PLRV RNA. Following RNase H and DNA polymerase I-mediated second strand synthesis, double-stranded cDNA was cloned into the Pst I site of the plasmid pUC9 using oligo (dC)-oligo(dG) tailing methodology. Escherichia coli JM109 transformants were screened with first-strand /sup 32/P-cDNA in colony hybridization experiments to confirm that recombinants contained PLRV-specific sequences.},

  doi          = {},

  url          = {https://www.osti.gov/biblio/7187998},
  journal      = {},
number       = ,

  volume       = ,

  place        = {United States},

  year         = {Thu Jan 01 00:00:00 EST 1987},

  month        = {Thu Jan 01 00:00:00 EST 1987}

}

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