MLSBIO101 RECOMBINANT DNA TECHNOLOGY Flashcards
What do you call DNA molecules that are extracted from different sources and chemically joined together?
Recombinant DNA Technology
What is the DNA comprising animal gene which may be recombined with DNA from a bacterium?
Recombinant DNA Technology
What year is the discovery of DNA structure by Watson and Crick?
1953
Who discovered DNA structure in 1953?
Watson and Crick
What year is the isolation of DNA ligase?
1967
What year is the isolation of REase?
1970
What year did Paul Berg generated rDNA technology?
1972
Who generated rDNA technology in 1972?
Paul Berg
What year did Cohen and Boyer produced the first plasmid vector capable of being replicated within a bacterial host?
1973
Who produced the first plasmid vector capable of being replicated within a bacterial host?
Cohen and Boyer
What are the goals of recombinant DNA technology?
- To isolate and characterize a gene
- TO make desired alterations in one or more isolated genes
- To return altered genes to living cells
- Artificially synthesize new gene
- Alternating the genome of an organism
- Understanding the hereditary diseases and their cure
- Improving human genome
What are the 4 procedures of making rDNA?
- Isolating of DNA
- Cutting of DNA
- Joining of DNA
- Amplifying of DNA
In Isolation of DNA, cells are lysed using ____ in order to disrupt the plasma membrane.
Detergent
In Isolation of DNA, after lysing the cells, it is treated with____ to destroy protein and ___ to destroy RNA.
Protease and RNAase
In isolation of DNA, after centrifuging the test tube, what do you transfer to a clean tube?
Supernatant
In Isolation of DNA, What do you use to precipitate the DNA after transferring the supernatant to a clean test tube?
Ethanol
In cutting of DNA, how does DNA cut into large fragments?
Mechanical Shearing
In cutting of DNA, what are the scissors of molecular genetics?
Restriction Enzymes
What is a special class of sequence-specific enzyme?
Restriction Enzyme
What enzyme is found in bacteria?
Restriction Enzyme
T/F. Site-specific-cleave DNA molecules only cleaves at specific nucleotide sequence.
True
What characteristic do REases recognize DNA base sequence?
Palindrome
T/F. REase make staggered cuts with complementary base sequences for easy circulization.
True
In amplifying the recombinant DNA, To what is the recombinant DNA transformed into?
Bacterial host strain
In Amplification, what do you treat the cells?
CaCl2 or Calcium chloride
In amplification, what temperature are the cells heat shocked?
42 C
What enzyme binds to DNA molecules?
DNA Ligase
What enzyme cleaves DNA at specific sites?
Type II Restriction Endonuclease
What enzyme makes a DNA copy of RNA molecule?
Reverse Transcriptase
What enzyme fills single stranded gapes of DNA duplex?
DNA Polymerase I
What enzyme adds phosphate to the 5’-OH end of a polynucleotide?
Polynucleotide Kinase
What enzyme adds homopolymer tails to the 3’-OH ends?
Terminal Transferase
What enzyme removes nucleotide residues from 3’ ends?
Exonuclease III
What enzyme removes nucleotides from the 5’ ends?
Bacteriophage {Lamda} Exonuclease
What enzyme removes terminal phosphates?
Alkaline Phosphatase
What do you call the area of DNA that can join another DNA part without losing the limit for self-replication?
Vector
T/F. A vector does not lose the limit for self-replication.
True
What is capable of replicating in host cell?
Vector
T/F. A vector should have convenient RE sites for inserting DNA of interest.
True
T/F. A vector should have a selectable marker to indicate which host cells received recombinant DNA molecule?
True
T/F. A vector should be small and easy to isolate.
True
What do you call the small, circular DNA molecules that are separated from the rest of the chromosome?
Plasmids
What vector replicates independently of the bacterial chromosome?
Plasmids Vector
What vector is useful for cloning DNA inserts less than 20 kb (kilobase pairs)?
Plasmids Vector
What vector is the recombinant infections, containing the phage chromosome in addition to embedded “outside” DNA?
Lamba Phage Vectors
T/F. Phage vectors can convey bigger DNA groupings than plasmid vectors.
True
What vector is the hybrids of phages and plasmids that can carry DNA fragments up to 45kb?
Cosmids
What vector can replicate like plasmids but can package like phage lambda?
Cosmids
What are the vectors that carry host signals that facilitate the transcription and translation of an inserted gene?
Expression Vectors
What vector is useful for expressing eukaryotic genes in bacteria?
Expression Vectors
What vector has been engineered to contain a centromere, telomere, origin of replication, and a selectable marker?
Yeast Artificial Chromosome (YACS)
What vector can carry up to 1000 kb of DNA?
Yeast Artificial Chromosomes (YACS)
What vector is useful for cloning eukaryotic genes that contains introns?
Yeast Artificial Chromosomes (YACS)
What vector are bacterial plasmids derived from the F plasmid?
Bacterial Artificial chromosomes (BACS)
What vector is capable of carrying up to 300kb of DNA?
Bacterial Artificial Chromosomes (BACS)
What are the techniques used in rDNA technology?
- Gel Electrophoresis
- Cloning Libraries
- Restriction Enzyme Mapping
- PCR
- Nucleic Acid Hybridization
- DNA Microarrays
What technique are DNA fragments of different sizes can be separated by an electrical field applied to a gel?
Gel Electrophoresis
What technique does the negatively charged DNA migrates away from the negative electrode and to the positive electrode?
Gel Electrophoresis
T/F. In gel electrophoresis, the smaller the fragment, the faster it migrates.
True
What do you call the collection of DNA clones in a certain vector?
Libraries
T/F. The goal of cloning libraries is to have each gene representing in the library at least once.
True
What is made from RE DNA fragments of total genomic DNA?
Genomic
What is made from DNA synthesized from mRNA?
cDNA (Complementary DNA)
What technique allows the isolation of specific segment of DNA from a small DNA (or cell sample) using DNA primers at the ends of the segment of interest?
PCR
T/F. It is important to have a restriction enzyme site map of a cloned gene for further manipulations of the gene.
True
What is accomplished by digestion of the gene singly with several enzymes and then in combinations?
Restriction Enzyme Mapping
T/F. In restriction enzyme mapping, the fragments are subjected to gel electrophoresis to separate the fragments by size and the sites are deduced based on the sizes of the fragments.
True
In Nucleic Acid Hybridization, what blot allows the detection of a gene of interest by probing DNA fragments that have been separated by electrophoresis with a “labeled” probe?
Southern Blot
In Nucleic Acid Hybridization, what blot probe RNA on a gel with DNA probe?
Northern Blot
In Nucleic Acid Hybridization, what blot probes proteins on a gel with an antibody?
Western Blot
What technique is the vast majority of the protein-encoding qualities onto a microarray chip, utilizing innovation in light of the DNA silicon chip industry?
DNA Microarrays
In DNA Microarrays, what is utilized to hybridize to cell RNA, and measure the statement rates of a substantial number of qualities in a cell?
Chip
To what are applies the growing crops of your choice (GM food), pesticide resistant crops, fruits with attractive colors, all being grown in artificial conditions?
Agriculture
To what area applies where artificial insulin production, drug delivery to target sites?
Pharmacology
To what area applies gene therapy, antiviral therapy, vaccination, and synthesizing clotting factors?
Medicine
