PART1(Postmidterm) Flashcards
molecule contains a combination of DNAs of different origins; combining of different DNAs is made possible by the use
restriction enzymes and ligase
recombinant DNA
Two categories of enzymes needed for recombinant DNA
Restriction endonucleases
DNA ligase
act as scissors that cut both strands of the DNA sugar-phosphate backbone; these enzymes are found primarily in bacteria
Restriction endonucleases
– joins two DNA molecules
DNA ligase
Two Types of ends found in DNA
strands after being cut by restriction
enzymes
Blunt ends
Sticky end
is when both strands are
of equal length and have no
unpaired bases.
Blunt end
is when one strand is longer than the other and has unpaired bases that can form hydrogen bonds with complementary
bases.
Sticky end
are helpful in cloning because they can be joined by DNA ligase.
Sticky ends
can then join DNA fragments with complementary sticky ends or
blunt ends
DNA ligase
catalyzes the formation of covalent bonds between the sugar and the phosphate of adjacent nucleotides,
Ligase
it is called _______ when the two different DNA fragments become one DNA molecule
Recombinant DNA
Molecular cloning is also known as
DNA cloning
- DNA molecules that are used as vehicles in transporting the foreign piece of DNA into the host cell;
Cloning vectors
Cloning victors types
Plasmid vectors
Cosmid Vectors
Phage DNA vectors
BAC vectors
YAC vectors
Plant Cloning vectors
is a popular and effective molecular biology tool often used to detect recombinant bacteria in cloning experiments.
Blue-White Screening
These are small, double strands of bacterial DNA that are extrachromosomal
Plasmid Vectors
are cloning vectors derived from bacteriophages, which are viruses that infect bacteria. These vectors are used to carry and replicate foreign DNA within a host/bacteral cell.
Phage DNA vectors
These are engineered hybrids of phage DNA and plasmid. it contains cos sites for packaging into phage particles, plasmid replication origin for replication in bacterial hosts and genes for plasmid selection
Cosmid vectors
This type of vector is useful for analyzing large portions of complex genome and it is an engineered DNA molecule (synthetic) used to clone DNA sequences in
bacterial cells that have been widely used in large genome sequencing projects
BAC vectors
These are useful for eukaryotic molecular studies and allow complete genes and gene clusters to be cloned for study. They are particularly useful for studying complex genomes and generating large-insert libraries
YAC vectors
A special region in the Ti plasmid, called T-DNA, containing about 8 genes causing the disease characteristics, is incorporated into the plant’s genome so that the plant will synthesize some nutrients (called opines) that the bacterial cells use.
Plant Cloning Vectors
can be used to transfer foreign genes into plants.
T-DNA
lacks some genes that cause the tumor.
Ti plasmid
