10.1.1 Recombinant DNA Technology

Recombinant DNA technology is the foundation of genetic engineering, which involves a series of steps to join DNA segments from different sources. This is an in-vivo (in living cells) method which is used for the synthesis of product of gene beside copies of desired gene (gene of interest) at industrial scale.

Definition

Recombinant DNA Technology: Recombinant DNA technology is the foundation of genetic engineering, which involves a series of steps to join DNA segments from different sources. This is an in-vivo (in living cells) method which is used for the synthesis of product of gene beside copies of desired gene (gene of interest) at industrial scale.

Main Idea

• Recombinant DNA technology is the foundation of genetic engineering.
• It involves a series of steps to join DNA segments from different sources.
• It is an in-vivo (in living cells) method.
• It is used for the synthesis of product of gene.
• It is also used for making copies of desired gene (gene of interest).
• It is used at industrial scale.
• It involves the selection and isolation of gene of interest.
• It involves inserting the gene of interest in a suitable vector.
• It involves the transformation into suitable host by the recombinant DNA (rDNA).
• Components or requirements of recombinant DNA technology include:

Core Concept

• Recombinant DNA technology = foundation of genetic engineering
• In-vivo method = in living cells
• Purpose = synthesis of product of gene; copies of desired gene
• Scale = industrial scale
• Steps = selection and isolation of gene of interest; inserting in vector; transformation into host
• Components = gene of interest; molecular scissors; molecular carrier or vector; molecular glue; expression system

Detailed Explanation

Recombinant DNA Technology

• Recombinant DNA technology is the foundation of genetic engineering, which involves a series of steps to join DNA segments from different sources.
• This is an in-vivo (in living cells) method which is used for the synthesis of product of gene beside copies of desired gene (gene of interest) at industrial scale.
• It involves the selection and isolation of gene of interest, inserting it in a suitable vector and the transformation into suitable host by the recombinant DNA (rDNA).
• Components or requirements of recombinant DNA technology include:

Gene of Interest

The gene of interest is the gene which is to be cloned. It can be obtained by one of the three possible ways:

a. Artificial gene synthesis is the process of synthesizing a gene in-vitro (Latin: "in glassware" means "outside the living cell") without template DNA samples with the help of DNA synthesizer machine.

b. Gene of interest can also be obtained by synthesizing it from its mRNA. Synthesis of gene from mRNA is carried out by reverse transcriptase enzyme which is naturally found in retroviruses. The DNA formed by this process is called complementary DNA (cDNA).

c. In most of the cases the gene of interest is directly cleaved from a chromosomal DNA by using particular DNA scissors called restriction endonucleases.

Summary Table of Components

Summary of Obtaining Gene of Interest

Intermediate Level

Recombinant DNA Technology

Components of Recombinant DNA Technology

Obtaining Gene of Interest

Formula / Mathematical Form

Not applicable to this topic.

Solved Example

Not applicable to this topic (descriptive topic).

Important Points

• 1. Recombinant DNA technology is foundation of genetic engineering.
• 2. It involves joining DNA segments from different sources.
• 3. It is an in-vivo method.
• 4. It is used for synthesis of product of gene.
• 5. It is used for making copies of desired gene.
• 6. It is used at industrial scale.
• 7. It involves selection and isolation of gene of interest.
• 8. It involves inserting gene of interest in a suitable vector.
• 9. It involves transformation into suitable host.
• 10. Components include gene of interest.
• 11. Components include molecular scissors.
• 12. Components include molecular carrier or vector.
• 13. Components include molecular glue.
• 14. Components include expression system.
• 15. Gene of interest can be obtained by artificial gene synthesis.
• 16. Gene of interest can be obtained from mRNA.
• 17. Gene of interest can be obtained from chromosomal DNA.
• 18. Artificial gene synthesis uses DNA synthesizer machine.
• 19. Synthesis from mRNA uses reverse transcriptase.
• 20. Cleaving from chromosomal DNA uses restriction endonucleases.

Remember

• - Recombinant DNA technology = foundation of genetic engineering
• - In-vivo method = in living cells
• - Purpose = synthesis of product of gene; copies of desired gene
• - Scale = industrial scale
• - Steps = selection and isolation of gene of interest; inserting in vector; transformation into host
• - Components = gene of interest; molecular scissors; molecular carrier or vector; molecular glue; expression system

Common Mistakes

• 1. Confusing recombinant DNA technology and genetic engineering — Recombinant DNA technology is the foundation of genetic engineering.
• 2. Forgetting the in-vivo nature — It is an in-vivo method.
• 3. Mixing up the components — Gene of interest, molecular scissors, vector, glue, expression system.
• 4. Forgetting the methods of obtaining gene of interest — Artificial synthesis, from mRNA, from chromosomal DNA.
• 5. Mixing up the enzymes — Reverse transcriptase for cDNA; restriction endonucleases for cleaving.
• 6. Believing recombinant DNA technology is only for medicine — It has many applications.
• 7. Mixing up the scale — Industrial scale.
• 8. Forgetting the purpose — Synthesis of product of gene and copies of desired gene.
• 9. Mixing up the steps — Selection, isolation, insertion, transformation.
• 10. Thinking recombinant DNA technology is simple — It involves many steps and components.

Quick Revision

Recombinant DNA technology is the foundation of genetic engineering, which involves a series of steps to join DNA segments from different sources. This is an in-vivo (in living cells) method which is used for the synthesis of product of gene beside copies of desired gene (gene of interest) at industrial scale. It involves the selection and isolation of gene of interest, inserting it in a suitable vector and the transformation into suitable host by the recombinant DNA (rDNA). Components or requirements of recombinant DNA technology include: 1. Gene of interest 2. Molecular scissors 3. Molecular carrier or vector 4. Molecular glue 5. Expression system. The gene of interest is the gene which is to be cloned. It can be obtained by one of the three possible ways: a. Artificial gene synthesis is the process of synthesizing a gene in-vitro (Latin: "in glassware" means "outside the living cell") without template DNA samples with the help of DNA synthesizer machine. b. Gene of interest can also be obtained by synthesizing it from its mRNA. Synthesis of gene from mRNA is carried out by reverse transcriptase enzyme which is naturally found in retroviruses. The DNA formed by this process is called complementary DNA (cDNA). c. In most of the cases the gene of interest is directly cleaved from a chromosomal DNA by using particular DNA scissors called restriction endonucleases.

60-Second Revision

• Recombinant DNA technology = foundation of genetic engineering
• In-vivo method = in living cells
• Purpose = synthesis of product of gene; copies of desired gene
• Scale = industrial scale
• Steps = selection and isolation of gene of interest; inserting in vector; transformation into host
• Components = gene of interest; molecular scissors; molecular carrier or vector; molecular glue; expression system
• Warning: Recombinant DNA technology is an in-vivo method

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• 10.2 MOLECULAR SCISSORS (RESTRICTION ENDONUCLEASE)

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