Which vectors can be used to clone recombinant DNA?
The most commonly used cloning vectors are E. coli plasmids, small circular DNA molecules that include three functional regions: (1) an origin of replication, (2) a drug-resistance gene, and (3) a region where DNA can be inserted without interfering with plasmid replication or expression of the drug-resistance gene.
How do you choose a vector for cloning?
Despite the bewildering choice of commercial and other available vectors, the choice of which cloning vector to use can be decided by applying a small number of criteria: insert size, copy number, incompatibility, selectable marker, cloning sites, and specialized vector functions.
Which is used as a cloning vector into plants?
Ti plasmid
How does a plasmid work as a cloning vector?
Plasmids are autonomously replicating circular extra-chromosomal DNA. They are the standard cloning vectors and the ones most commonly used. Most general plasmids may be used to clone DNA insert of up to 15 kb in size. One of the earliest commonly used cloning vectors is the pBR322 plasmid.
What are the features of a good cloning vector?
- it must be small in size.
- It must be self-replicating inside host cell.
- It must possess restriction site for Restriction Endonuclease enzymes.
- Introduction of donor DNA fragment must not interfere with replication property of the vector.
What are the three features of cloning vector?
Characteristics of a cloning vectors
- it must be small in size.
- It must be self-replicating inside host cell.
- It must possess restriction site for Restriction Endonuclease enzymes.
- Introduction of donor DNA fragment must not interfere with replication property of the vector.
What are the 4 major components of a cloning vector that you need in order to clone?
components of plasmid cloning vectors:
- origin of replication (ori) site where DNA replication is initiated.
- marker genes for selection and/or screening.
- Unique restriction endonuclease (RE) sites. – allow inserts to be cloned in specific sites on plasmid.
- transmissability.
- Promoters for gene expression.
What are cloning vectors used for?
In general, cloning vectors are plasmids that are used primarily to propagate DNA. They replicate in E. coli to high copy numbers and contain a multiple cloning site (also called a polylinker) with restriction sites used for inserting a DNA fragment.
What other vectors can be used?
The four major types of vectors are plasmids, viral vectors, cosmids, and artificial chromosomes. Of these, the most commonly used vectors are plasmids. Common to all engineered vectors have an origin of replication, a multicloning site, and a selectable marker.
Which vector has highest capacity?
Vector size: The vector size range varies from 5 kb plasmid vectors to 6–10 megabases HAC high-capacity vectors….1.1 Introduction.
| Vector | Host | Insert size |
|---|---|---|
| PACs | E.coli | 100–300kb |
| BACs | E.coli | <300 kb |
| YACs | Saccharomyces cerevisae | 200–2000kb |
| Human Artificial Chromosomes (HACs) | Cultured Human Cells | >2000kb |