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Cloning of herpes simplex type 1 DNA fragments in a bacteriophage lambda vector
Science (New York, N.Y.)
|February 9, 1979
Summary
Researchers created hybrid DNA by inserting herpes simplex type 1 (HSV-1) DNA fragments into a bacteriophage vector. Analysis revealed DNA from approximately 50% of the HSV-1 genome and identified distinct classes of defective HSV-1 genomes.
Area of Science:
- Molecular Biology
- Virology
- Genetics
Background:
- Herpes simplex type 1 (HSV-1) is a common human pathogen.
- Understanding the HSV-1 genome, including defective variants, is crucial for virology and disease research.
Purpose of the Study:
- To construct and analyze recombinant DNA hybrids between HSV-1 and a bacteriophage vector.
- To investigate the genetic makeup of both normal and defective HSV-1 virions.
Main Methods:
- DNA isolation from HSV-1 virions (defective and nondefective).
- Restriction endonuclease digestion of HSV-1 DNA.
- Insertion of DNA fragments into the EK2 coliphage lambda gtWES . lambda B vector.
- In vitro encapsidation and purification of lambda-HSV1 hybrids under P4 containment.
- Analysis of isolated recombinant DNA.
Main Results:
- Successfully created lambda-HSV1 DNA hybrids containing fragments from approximately 50% of the normal HSV-1 genome.
- Identified multiple, distinct classes of defective HSV-1 genomes based on the analysis of hybrids derived from defective virions.
- Demonstrated the feasibility of using bacteriophage vectors for cloning and analyzing viral DNA under stringent containment.
Conclusions:
- The study provides a method for analyzing complex viral genomes using recombinant DNA technology.
- The findings confirm the heterogeneity of defective HSV-1 genomes, suggesting multiple origins or mechanisms of defectiveness.
- This approach facilitates further genetic characterization of HSV-1 and its variants.