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  Section: General Biotechnology / Genes & Genetic Engineering
 
 
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Genes : Nature, Concept and Synthesis

 
     
 

Gene Expression
The DNA has two important roles in the cell, first is replication and the second is expression. Gene expression accomplished by a series of events that contained in DNA is converted into molecule that take place in the cell. The information contained in DNA is converted into molecules that determines the metabolism of the cell. During the process of gene expression DNA is first copied into an mRNA molecule which determines the amino acid sequence of a molecule of protein. The RNA molecules are synthesized by using a portion of base sequences of single strand of double stranded DNA. This single strand is called template. Hence formation of an RNA transcript is facilitated by an enzyme RNA polymerase. Therefore, the process of synthesis of an RNA molecule corresponding to a gene is called transcription. By using base sequences and RNA molecule proteins are synthesized in a definite order. Production of an amino acid Sequence from an mRNA base sequence is called translation. After completion of translation proteins are synthesized. Therefore gene expression refers to proteins synthesis through two major events, transcription and translation.

Central Dogma
DNA itself cannot directly order for the synthesis of amino acids but forms its transcripts first which is then translated into protein. For the first time in 1958, F. Crick suggested that there is unidirection flow of informations from DNA to RNA to protein shown as below:

DNA

Transcription

RNA

Translation

Protein


This sequential transfer of information from DNA to protein via RNA is known as central dogma. Further more in 1963 H.M. Temin and coworkers reported that Rous sarcoma virus causing cancer contains RNA as genetic material. In1964, he put forward a hypothesis that RNA tumour viruses synthesize an enzyme reverse transcriptase that synthesized DNA from RNA template. It could also be demonstrated that DNA synthesis prevented after destroying enzyme RNase. It was also shown that RNA specific DNA was synthesized. However, in lymphocytes of leukemia patients RNA-dependent DNA polymerase was discovered. In 1970 Crick again suggested a modified version of central dogma (Fig.2.10)
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Content

Chemical nature of DNA

 

Chemical composition

 

Nucleotides, nucleosides

 

Polynucleotides

 

Chargaff's rule of equivalence

Physical nature of DNA

 

Watson and Cricks model of DNA

 

Circular and superhelical DNA

 

Organization of DNA in eukaryotes

Structure of RNA

Gene concept

Units of a gene

 

Cistron

 

Recon

 

Mutan

Split genes (introns)

 

RNA splicing

 

Ribozyme

 

Evolution of split genes

Overlapping gene

Gene organization

Gene expression

Gene regulation

 

Transcription

 

 

The lac operon (structural gene, operator gene, promoter gene and repressor gene)

Artificial synthesis of genes

 

Synthesis of a gene for yeast alanine tRNA

 

Synthesis of a gene for bacterial tyrosine tRNA

 

Synthesis of a human leukocyte interferon gene

Gene synthesis by using mRNA

Gene machine

The PCR

 

Amplification of DNA (melting of target DNA, annealing of primers, primer extension)

 

Application of PCR technology

   

DNA can undergo replication process to form DNA and transcription to form RNA that in turn undergoes translation. RNA also replicates to form RNA. . In special cells only (dot lines) RNA synthesizes DNA, and DNA synthesizes proteins.

Modified central dogma i.e. RNA dependent DNA synthesis  
This process is known as RNA directed DNA synthesis (Temin, 1972). For detailed discussion of transcription and translation see A Text Book of Microbiology by Dubey and Maheshwari (1999).

Fig 2.10. Modified central dogma i.e. RNA dependent DNA synthesis

   
 
     
 
 
     



     
 
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