Generation of a plasmid vector for deletion cloning by rapid multiple site-directed mutagenesis
| Autor: | Kolari S. Bhat |
|---|---|
| Rok vydání: | 1993 |
| Předmět: |
Genetics
Base Sequence Genetic Vectors Molecular Sequence Data Cloning vector General Medicine DNA Biology Molecular cloning Restriction enzyme Restriction site Subcloning Plasmid Mutagenesis Site-Directed Cloning Molecular Site-directed mutagenesis Deoxyribonucleases Type II Site-Specific T-DNA Binary system Plasmids Sequence Deletion |
| Zdroj: | Gene. 134(1) |
| ISSN: | 0378-1119 |
| Popis: | The construction of a new plasmid vector, devoid of all Mbo I (GATC) and Tsp EI (AATT) restriction sites, is described. The lack of these two frequent-cutting restriction sites is a unique feature among plasmids. This new plasmid, pBRkanf1 − , allows selective fragmentation of a cloned insert. As a result, the vector offers an alternative strategy to create overlapping and sequentially deleted subclones. In addition, the construction of the new plasmid required the development of a rapid and accurate multiple site-directed mutagenesis procedure. The mutagenesis method uses a combination of DNA amplification and chain extension by DNA polymerase. By this method, mutations are created progressively from one end of a DNA molecule to the other. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|