Efficient generation of infectious recombinant baculoviruses by site-specific transposon-mediated insertion of foreign genes into a baculovirus genome propagated in Escherichia coli
| Autor: | G F Barry, Verne A. Luckow, Stephen C. Lee, P O Olins |
|---|---|
| Rok vydání: | 1993 |
| Předmět: |
Transposable element
viruses Immunology Genetic Vectors Molecular Sequence Data Restriction Mapping BacMam Genome Viral Biology Moths medicine.disease_cause Recombinant virus Transfection Microbiology Polymerase Chain Reaction Cell Line Plasmid Transformation Genetic Shuttle vector Virology medicine Escherichia coli Animals Replicon Cloning Molecular Recombination Genetic Base Sequence fungi food and beverages Molecular biology Transformation (genetics) Mutagenesis Insertional Oligodeoxyribonucleotides Insect Science DNA Transposable Elements Mutagenesis Site-Directed Baculoviridae Plasmids Research Article |
| Zdroj: | Journal of virology. 67(8) |
| ISSN: | 0022-538X |
| Popis: | The construction and purification of recombinant baculovirus vectors for the expression of foreign genes in insect cells by standard transfection and plaque assay methods can take as long as 4 to 6 weeks. This period can be reduced to several days by using a novel baculovirus shuttle vector (bacmid) that can replicate in Escherichia coli as a plasmid and can infect susceptible lepidopteran insect cells. The bacmid is a recombinant virus that contains a mini-F replicon, a kanamycin resistance marker, and attTn7, the target site for the bacterial transposon Tn7. Expression cassettes comprising a baculovirus promoter driving expression of a foreign gene that is flanked by the left and right ends of Tn7 can transpose to the target bacmid in E. coli when Tn7 transposition functions are provided in trans by a helper plasmid. The foreign gene is expressed when the resulting composite bacmid is introduced into insect cells. |
| Databáze: | OpenAIRE |
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