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Group 5 Enzyme Vector Used in Rdna Technology

The document provides an overview of enzymes and vectors used in recombinant DNA technology. Key enzymes include DNA ligase, restriction endonucleases, and reverse transcriptase, while vectors discussed include plasmids, lambda phage, cosmids, expression vectors, bacterial artificial chromosomes (BACs), and yeast artificial chromosomes (YACs). Each vector type has unique characteristics and capacities for DNA insertion, making them suitable for various cloning and expression applications.

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70 views10 pages

Group 5 Enzyme Vector Used in Rdna Technology

The document provides an overview of enzymes and vectors used in recombinant DNA technology. Key enzymes include DNA ligase, restriction endonucleases, and reverse transcriptase, while vectors discussed include plasmids, lambda phage, cosmids, expression vectors, bacterial artificial chromosomes (BACs), and yeast artificial chromosomes (YACs). Each vector type has unique characteristics and capacities for DNA insertion, making them suitable for various cloning and expression applications.

Uploaded by

ishikawareikosan
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
Available Formats
Download as PPTX, PDF, TXT or read online on Scribd

GROUP 5

10 minutes Presentation
Enzymes used in recombinant DNA
technology

DNA ligase • Bind to DNA molecules

Type II restriction endonuclease • Cleaves DNA at specific sites


Reverse transcriptase • Make a DNA copy of RNA molecule

DNA polymerase I • Fill single stranded gapes of DNA duplex

Polynycleotide Kinase • Adds a phosephate to the 5'-OH end of a


polynucleotide
Terminal transferase • Adds homopolymer tails to the 3'-OH ends

Exonuclease III • Removes nucleotide residues from the 3' ends


Bacteriophage {lamda} • removes nucleotides from the 5' ends
exonuclease
Alkaline phosphatase • Removes terminal phosphates
Vectors used in rDNA technology

• A vector is an area of DNA that can join


another DNA part without losing the limit for
self-replication
• Should be capable of replicating in host cell
• Should have convenient RE sites for
inserting DNA of interest
• Should have a selectable marker to indicate
which host cells received recombinant DNA
molecule
Vectors used in rDNA technology

BAC
S
plasm YAC
id S
vector
s
Lamd
expre
a
ssion
phage
cosmi
d
Plasmid vector

• Plasmids are small, circular DNA


molecules that are separate from the
rest of the chromosome.
• They replicate independently of the
bacterial chromosome.
• Useful for cloning DNA inserts less
that 20 kb (kilobase pairs).
• Inserts larger than 20 kb are lost
easily in the bacterial cell.
Lamda phage vector

• Lamda phage vectors are recombinant


infections, containing the phage
chromosome in addition to embedded
"outside" DNA.
• All in all, phage vectors can convey
bigger DNA groupings than plasmid
vectors.
Cosmid vector

• Cosmids are hybrids of


phages and plasmids
that can carry DNA
fragments up to 45 kb.
• They can replicate like
plasmids but can be
packaged like phage
lambda
Expression vectors

• Expression vectors are


vectors that carry host
signals that facilitate
the transcription and
translation of an
inserted gene.
• They are very useful
for expressing
eukaryotic genes in
bacteria.
Yeast artificial chromosomes
(YACS)
• Yeast artificial chromosomes (YACS)
are yeast vectors that have been
engineered to contain a centromere,
telomere, origin of replication, and a
selectable marker.
• They can carry up to 1,000 kb of DNA.
• they are useful for cloning eukaryotic
genes that contain introns.
Bacterial artificial chromosomes
(BACS
• Bacterial artificial
chromosomes (BACS)
are bacterial plasmids
derived from the F
plasmid. They are
capable of carrying up
to 300 kb of DNA.

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