Help eplant science to add more and more information day by date and keep it updated
Help eplant science to add more and more information day by date and keep it updated

MoonHeart Infotech offers Web Designing, web development, SEO, Internet Marketing in Mumbai, Pune, Maharahtra at very cheap rates
 
 
Section: General Biotechnology / Genes & Genetic Engineering
 
     
 
Techniques of Genetic Engineering
 
   
 

Animal viruses
A variety of animal viruses are known, for example, simian virus 40 (SV40), adinovirus, retrovirus (ssRNA), vaccinia virus, etc. They can increase the efficiency of animal cell transformation if they are used as vector. A special feature of viruses is that they contain the strong promoters which can possibly bring about expression of inserted DNA fragment of the viruses. The most commonly used virus is SV40 which contains a circular DNA of about 5.2 Kb. In addition to ori region, the DNA contains early genes and late genes. Early genes are required for DNA replication, whereas late genes encode coat protein of the virus.

Like phage λ, the amount of DNA to be packed into virus capsid is limited. The genome has 3 identified non-essential regions; hence the use of non-defective viral vectors is limited. The defective virus vectors have missing genes whose infection process has to be helped by viral DNA constituting functional copies of missing genes. If the cos cells (the host monkey cells) are used as host, they can be transformed by a plasmid carrying the early region of SV40 with a defective replication origin, cos cells express wild type large T antigen and can support the growth of SV40. Recombinants containing foreign DNA inserted into early region can be propagated without the need of helper virus (Glover, 1984). In this way, the recombinant and helper can be separately recovered from the host.

 

Content

Ä Gene cloning in prokaryotes

 

Ä Isolation of DNA to be cloned 

 

Ä Insertion of DNA fragment into vector 

 

 

Ä Use of restriction Linkers

 

 

Ä Use of homopolymer tails

 

Ä Transfer of recombinant DNA into bacterial cells

 

Ä Selection of clones

 

 

Ä Colony hybridization techniques

 

 

Ä In vitro translation technique

 

 

Ä Immunological tests

 

 

Ä Blotting Techniques

 

Ä Recovery of cells

 

Ä Expression of cloned DNA

 

 

Ä Shine-Dalgano sequence

 

 

Ä Expression vectors

Ä Gene cloning in eukaryotes

 

Ä Plant cells

 

 

Ä Yeasts

 

 

Ä Filamentous fungi

 

 

Ä Agrobacterium plasmids

 

 

Ä Plant cell transformation

 

 

Ä Plant cell transformation by ultrasonication

 

 

Ä Liposome mediated gene transfer

 

Ä Animal cell  

 

 

Ä Animal viruses

 

 

Ä Electroporation

 

 

Ä Particle bombardment

 

 

Ä Microinjection

 

 

Ä Direct transformation

Ä Site directed mutagenesis

 

Ä Methods of mutagenesis

 
     
  Go To Top  
     
  Created & Maintained by MoonHeart Infotech Copyright © 2009 ePlantScience.com