Publish Your Research Online
Get Recognition - International Audience
Request for an Author Account | Login | Submit Article
|HOME||FAQ||TOP AUTHORS||FORUMS||PUBLISH ARTICLE|
Gene Cloning in Plasmid VectorBY: Dr. Suresh Kaushik | Category: DNA | Submitted: 2013-07-04 10:51:56
Article Summary: "The terms recombinant DNA technology, DNA cloning, molecular cloning, or gene cloning all refer to the same process: the transfer of a DNA fragment of interest from one organism to a self-replicating genetic element such as a bacterial plasmid (cloning vector). The DNA of interest can then be propagated in a foreign host cell. T.."
Gene Cloning in Plasmid Vector | By - Dr. Suresh Kaushik
Gene cloning is a technique which is commonly used in biotechnology. In this technique a section of DNA is put into a vector that acts as a vehicle to transfer the DNA to a host cell e.g. a bacterium. The plamid vector multiplies in the bacterial cell. Thus, many copies of the original section of DNA are produced. Ultimately, a colony of bacteria forms, each containing one or several copies of the section of DNA contained in the vector DNA. The colony is termed a clone and the molecule of DNA contained in the vectors has been cloned. If the original section of DNA represented a gene, the process is known as gene cloning.
A DNA fragment to be cloned is obtained through the application of restriction endonucleases. Most of restriction enzymes cleave duplex DNA at specific palindromic sequences. An inherent advantage of this method is the ability to precisely excise the desired fragment through its restriction sites. The complimentary ends of the two DNAs specifically are covalently joined through action of the enzyme DNA ligase to form recombinant plasmid.
How can one select only those host organisms that contain a properly constructed vector? In the case of plasmid transformation, this is usually done through the use of antibiotics and/or chromogenic substrates. Addition of an antibiotic such as ampicillin will eliminate any colonies that did not take up the plasmid. The plasmids intact gene contains a gene that confers antibiotic resistance. In case of blue/white colony selection, E. coli transformed with a plasmid containing a foreign DNA insert in its polylinker region that lacks β-galactosidase activity because the insert interrupts the protein-encoding sequence of the lacZ' gene. Thus, when grown in the presence of blue dye X-gal bacterial colonies that have an insert in their polylinker region form colorless colonies, whereas bacteria containing only plasmids that lack an insert form blue colonies.
A cloned structural gene must be inserted into an expression vector, a plasmid that contains the properly positioned transcriptional and translational control sequences for the protein's expression. With the use of a relaxed control plasmid and an efficient promoter, the production of protein of interest may reach thirty percent of the host's total cellular protein. The ability to synthesize a given protein in large quantities is already having enormous medical, agricultural, and industrial impact.
Gene cloning has contributed hugely to agricultural and medical research. It has mainly been applied in agriculture to develop genetically modified crops or transgenic crops resistance to pests and diseases and improve nutritional quality of crops. It has allowed genes and their functions to be studied in much greater detail than was previously possible. In the future, there will be further possibilities for understanding gene function, protein formation and metabolism using gene cloning techniques.
About Author / Additional Info:
Dr. Suresh Kaushik
A Biotechnogical Professional and author from India
Comments on this article: (0 comments so far)
• UPR and its Consequences in Protein Maturation
• Laboratory Data For Blood Test
• Bioinformaticist vs. Bioinformatician - Definition, Differences and Career Outlook
• Proteomics and Analysis of Protein Sequence
Latest Articles in "DNA" category:
• Identifying a Specific Clone in CDNA and Genomic Library
• Biotechnolgical Techniques For DNA Analysis
• DNA Extraction:Procedure and Importance in Forensics
• Chromosomal Aberrations and its Types
• Gene Knockout in Mice
• DNA Repair Types: Excision, Postreplication, Recombination and Lesion Removal
• Microarrays and Gene Expressions - Principle and Procedure
• Human Cytogenetics - Karyotype
• Experimental Issues in Microarrays
• Nuclear pre-mRna Splicing: The Story of Introns and Exons
• Chain Termination Method: A Generic Method For DNA Sequencing
• Transposable Elements - The Story of Jumping Genes
• RNA Interference - The Art of Gene Silencing
• Protein Biosynthesis: Decoding the Code (Part-1)
• Protein Biosynthesis: Decoding the Code (Part - 2)
• Mutagenesis - Types and Uses
• C-Value, An Unsolved Paradox?
• Mechanism of Epigenetics
• Techniques of Epigenetic Studies
Important Disclaimer: All articles on this website are for general information only and is not a professional or experts advice. We do not own any responsibility for correctness or authenticity of the information presented in this article, or any loss or injury resulting from it. We do not endorse these articles, we are neither affiliated with the authors of these articles nor responsible for their content. Please see our disclaimer section for complete terms.
Copyright © 2010 biotecharticles.com - Do not copy articles from this website.
ARTICLE CATEGORIES :
| Disclaimer/Privacy/TOS | Submission Guidelines | Contact Us