What is the role of a polylinker or multi-cloning site in a plasmid used for cloning in E coli?Group of answer choicesSo that the inserted gene can be expressedSo a restriction enzyme can cut the plasmid in a specific locationSo that only the E coli cells that take up the plasmid will survive.So that the plasmid is maintained in the E coli cells as they grow and divide.
Question
What is the role of a polylinker or multi-cloning site in a plasmid used for cloning in E coli?Group of answer choicesSo that the inserted gene can be expressedSo a restriction enzyme can cut the plasmid in a specific locationSo that only the E coli cells that take up the plasmid will survive.So that the plasmid is maintained in the E coli cells as they grow and divide.
Solution
The role of a polylinker or multi-cloning site in a plasmid used for cloning in E. coli is to provide a specific location where a restriction enzyme can cut the plasmid. This is crucial for the insertion of the gene of interest.
Here's a step-by-step explanation:
-
The polylinker or multi-cloning site is a short segment of DNA which contains many (hence 'multi') restriction sites. These are specific sequences of DNA which restriction enzymes can recognize and cut.
-
When a gene of interest is prepared for cloning, it is also cut with restriction enzymes. The aim is to have the ends of the gene and the plasmid compatible for ligation.
-
The cut plasmid and the gene of interest are mixed together with a ligase enzyme, which facilitates the joining of the two DNA molecules.
-
The plasmid, now containing the gene of interest, is introduced into E. coli cells in a process called transformation.
-
The E. coli cells are then allowed to grow and divide, producing many copies of the plasmid and the gene of interest.
So, the polylinker or multi-cloning site does not directly contribute to the expression of the inserted gene, the survival of the E. coli cells that take up the plasmid, or the maintenance of the plasmid in the E. coli cells as they grow and divide. Its primary role is to provide a site for the restriction enzyme to cut the plasmid in a specific location, allowing for the insertion of the gene of interest.
Similar Questions
What is the role of the selectable marker in a plasmid for cloning in E coli? Group of answer choicesSo that the inserted gene can be expressedSo that the plasmid is maintained in the E coli cells as they grow and divide.So that only the E coli cells that take up the plasmid will survive.So a restriction enzyme can cut the plasmid in a specific location
Plasmids can be useful for many different elements of molecular microbiology but there are many considerations that dictate the features of a plasmid being used. If using a plasmid in order to express a large quantity of a particular protein (e.g. for protein purification) which of the following is NOT a requirement?The ability of the host bacterium to express GFP.A selectable marker that will facilitate retention of the plasmid.The presence of rep/ori genes that allows for replication of the plasmid in the bacterium of interest.A strong promoter that will drive transcription of the gene of interest and subsequent protein production.
Discuss the role of plasmid in working of bacteria.
Which of the following describes plasmids?Synthesise proteins.Controls what enters and exits the cell.Small circles of DNA that are used to share useful genes between bacteria.Contains all the instructions the bacterium needs to function and develop.
They are used in the process of insertion of genes into plasmid vectors during gene cloning and protein expression experiments. Select one: a. Restriction enzymes b. Terminal Transferase c. DNA polymerase d. Ligase
Upgrade your grade with Knowee
Get personalized homework help. Review tough concepts in more detail, or go deeper into your topic by exploring other relevant questions.