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Section: Genetics » Genetic Engineering and Biotechnology » Recombinant DNA and PCR
 
 
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  Artificial chromosome vectors for cloning large DNA segments
 
     
 
Content
⇒ Genetic Engineering and Biotechnology 1.  Recombinant DNA and PCR (Cloning and Amplification of DNA)
⇒ Restriction enzymes in cloning
⇒ Techniques used in recombinant DNA 
⇒ Cloning vectors for recombinant DNA
⇒ Plasmids as vectors
⇒ Bacteriophages as vectors
⇒ Plant and animal viruses as vectors
⇒ Transposons as vectors
⇒ Artificial chromosome vectors for cloning large DNA segments
⇒ Construction of chimeric DNA
⇒ Palindromes and staggered cleavage
⇒ Adding poly dA at the 3' ends of the vector and poly dT at the 3' ends of DNA clone
⇒ Blunt end ligation by T4 DNA ligase
⇒ Cloning in bacteria and eukaryotes
⇒ Cloning in bacteria
⇒ Cloning in eukaryotes
⇒ Molecular probes 
⇒ Labelling of probes
⇒ Applications of molecular probes
⇒ Construction and screening of genomic and cDNA libraries
⇒ Gene amplification : PCR and its applications
⇒ cDNA library from mRNA
⇒ Colony (or plaque) hybridization for screening of libraries
⇒ Gene Amplification : PCR and Its Applications
⇒ The basic polymerase chain reaction (PCR)
⇒ Different schemes of PCR


Artificial chromosome vectors for cloning large DNA segments
We know that in plasmids, sequences upto 10-15kb; in lambda (λ)phage, sequences upto 22kb and in cosmids sequences upto 40kb can be cloned. Yeast artificial chromosome vectors (YACs), allow cloning of sequences that are several hundred kilobase pairs long. These long molecules of DNA which may rarely represent whole chromosomes can be cloned in yeast by ligating them to vector sequences that allow their propagation as linear artificial chromosomes. These long DNA molecules can be generated and allow construction of comprehensive libraries (with large DNA segments) in microbial hosts. These vectors along with other yeast plasmid vectors are shown in Figure 39.11.
  
Five different yeast plasmid vectors.
Fig. 39.11. Five different yeast plasmid vectors.

 
     






     
     
 
 
     
 
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