What gene did the plasmid pSC101 carry?
pSC101 was the first cloning vector, used in 1973 by Herbert Boyer and Stanley Norman Cohen. Using this plasmid they have demonstrated that a gene from a frog could be transferred into bacterial cells and then expressed by the bacterial cells. The plasmid is a natural plasmid from Salmonella typhimurium.
What is the p15A origin?
Replication of Escherichia coli plasmid p15A was examined by use of a cell extract or a mixture of three purified E. Primer transcription starts 508 bp upstream of the replication origin. The region between 294 and 524 bp upstream of the origin determines the incompatibility property.
Why does a plasmid need an origin of replication?
The origin of replication also determines the plasmid’s compatibility: its ability to replicate in conjunction with another plasmid within the same bacterial cell. Plasmids that utilize the same replication system cannot co-exist in the same bacterial cell.
What does Pbr322 stand for?
pBR322 is a plasmid and was one of the first widely used E. coli cloning vectors. The p stands for “plasmid,” and BR for “Bolivar” and “Rodriguez, the scientists who synthesized the plasmid. So, the correct answer is ‘Bollivar and Rodrigues’
How many sets of antibiotic resistance does the plasmid Pbr322 carry?
How many sets of antibiotic resistance does the plasmid Pbr322 carry? Explanation: The plasmid contains two sets of antibiotic resistance genes on coding for the ampicillin resistance and the other for tetracycline resistance.
What is the origin of replication on the plasmid and why is it important to the genetic engineering process?
An origin of replication is the place where the process of DNA replication begins. It is a critical component of a DNA plasmid because it ensures the plasmid is passed from mother to daughter cells during cell division.
How the plasmid clones can be screened?
2. How the plasmid clones can be screened? Explanation: Plasmid contains a selectable marker that allows cells that contain the vector to be easily identified. Thus selectable markers are used to screen clones.
What is the temperature at which bacteria can take up the plasmid?
What is the temperature at which bacteria can takes up the plasmid? Explanation: Bacteria efficiently take up the plasmid DNA at -42˚C. This increases cell membrane permeability to DNA.
What is plasmid stability?
Structural plasmid stability exists, when all generated plasmids have the correct base sequence. When all daughter cells get at least one plasmid during cell division, the culture is segregational stable. The development of plasmid free cells can lead to a significant loss in productivity.
When is pSC101 excised from the host chromosome?
When shifted to the permissive temperature, the pSC101 replication origin in the host chromosome is detrimental, and the vector is excised from the chromosome.
Is the pSC101 replicon compatible with Pola function?
They are based on the pSC101 replicon consisting merely of the replication origin and repA gene, compatible with ColE1/pMB1/p15-derived plasmids, and thus do not require polA function of host cells. The copy numbers of the ts-Rep, tr and ts-Seg plasmids were 14, 5 and 1 per chromosome at 30 degrees C, respectively.
What’s the temperature at which plasmids are stable?
These plasmids are fairly stable when inherited at 30 degrees C, but not above 37 degrees C or 41.5 degrees C, depending on the repA mutations and host strains.