recombinant DNA technology Flashcards
a description of a clone
a technique scientists use to make exact genetic copies of living things. Genes, cells, tissues, and even whole animals can all be cloned. Some clones already exist in nature. Single-celled organisms like bacteria make exact copies of themselves each time they reproduce.
The first step in cloning a gene is to
b. Isolate the DNA from the organism that contains the desired gene
- Plasmids are put into bacterial cells by
d. Transformation
- Restriction enzymes
c. Make staggered cuts at specific sequences in DNA in both donor DNA and plasmid
- DNA ligase enzyme joins the cohesive ends of the desired DNA fragment with the cohesive ends of the plasmid
a. True
- Which of the following statements about the process of cloning is not true
valence. ……………..
d. Cloning produces a completely identical copy of an organism.
- Select the wrong statement about plasmids ?
a. It is extrachromosomal
b. It is double stranded
c. Its replication depends upon host cell
d. It is closed and circular DNA
valence……………………
Its replication depends upon host cell
- Conjugative plasmids
c. Carry transfer genes called the tra genes
note conjugative plasmid is a plasmid that is transferred from one bacterial cell to another during conjugation.
- Differentiate between plasmid and vector
- Plasmid is a small circular extrachromosomal DNA structure that replicates independently from chromosomal DNA in bacteria that carries foreign DNA molecules while
- Vector is a DNA fragment that carries foreign DNA molecules into the host cell for cloning purposes. E.g: plasmid, cosmid, bacteriophage, etc
Define “Cloning”
Cloning is a technique used to create exact genetic copies of genes, cells, or animals.
State three types of cloning
- Gene cloning: Involves creating copies of genes or DNA segments.
- Reproductive cloning: Involves creating copies of the whole organism.
- Therapeutic cloning: Involve in creation of embryonic stem cells.
Host cells that contain recombinant DNA are called……………
transformed cells
Some recombinant plasmids containing human DNA are inserted back into the bacterial cell (host cell) by the process called …………………………..
transformation
process of producing many copies of the gene of interest in the transformed cell through culture is called…………
amplification
what is a genomic library
is the collection of total genomic DNA from a single organism, the DNA is stored in the population of identical vectors, each containing different insert DNA.
A clone carrying the gene of interest can be identified with……………………… having a sequence complementary to the gene; the process called ………………………….
- nucleic acid probe
2. nucleic acid hybridization
what is a probe and what its use?
a probe is a nucleic acid that is complementary to the gene of interest.
it is used in the screen inorder tp identify the gene of interest in the genomic library.
difference between blunt end and sticky end?
blunt end are made by restriction enzymes where it leaves no unpaired bases on each end while sticky end are made by restriction enzymes so that it leaves unpaired ( 2 to 4 bases ) bases at each end of DNA.
what bond does restriction enzymes cut?
phosphodiester bond
DNA fragment acted upon by the same restriction enzyme has ………… sticky ends
complementary
enzymes that seal two complementary sticky ends are called
DNA ligase
complementary DNA/ cDNA
is is made by cloning the DNA made in reverse transcription of all mRNA produced in the cell.
polymerase chain reaction
is process of producing many copy of target DNA using taq polymerase
steps of PCR.
- denaturing, heat the target DNA at 94 degree Celsius for 1 min to unwind DNA to produce single strand of DNA,.
- annealing, mix single strands with primers and cool to 45 degree Celsius for 1 min to anneal.
- extension: add taq polymerase and raise temperature to 72 degree Celsius for 1 min. to get new double stranded DNA.
on double stranded DNA produce, two double stranded DNA in short period of time.
application of PCR.
- Amplification of DNA
