This chapter discusses the essential techniques and concepts of gene cloning in biotechnology. It explains how genes can be isolated and replicated for various applications, highlighting its significance.
Structured practice
Gene Cloning - Flash Cards
These flash cards cover important concepts from Gene Cloning in Biotechnology for Class 12 (Biotechnology).
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What is Gene Cloning?
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Gene cloning is the process of making multiple copies of a specific DNA fragment containing a gene of interest.
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What is a vector in gene cloning?
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A vector is a DNA molecule used to carry foreign genetic material into a host cell to facilitate cloning.
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3/19
Define recombinant DNA (rDNA).
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Recombinant DNA (rDNA) is formed by combining DNA from different sources, creating a DNA molecule that is not found naturally.
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What role do restriction enzymes play?
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Restriction enzymes, or restriction endonucleases, cut DNA at specific sequences, allowing for the insertion of foreign DNA.
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What is PCR?
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Polymerase Chain Reaction (PCR) is a technique used to amplify specific DNA sequences, making millions of copies for analysis.
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Explain the term 'host organism'.
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A host organism is a living cell that is used to introduce and replicate cloned DNA or recombinant DNA.
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What is the purpose of DNA ligase?
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DNA ligase is an enzyme that joins DNA fragments by forming phosphodiester bonds, essential for stabilizing recombinant DNA.
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What are plasmids?
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Plasmids are small, circular DNA molecules found in bacteria that can replicate independently and are often used as vectors in gene cloning.
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Define transformation in genetic engineering.
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Transformation is the process of introducing foreign DNA into a host cell, resulting in a genetically modified organism.
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What challenges are faced in gene isolation?
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Challenges include the vast genomic size, low concentration of the target gene, and susceptibility of DNA to physical or chemical degradation.
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What is the function of reverse transcriptase?
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Reverse transcriptase is an enzyme that synthesizes cDNA from an RNA template, crucial for cloning genes expressed as mRNA.
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Why is ETDA used during DNA extraction?
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EDTA chelates divalent cations, preventing the action of nucleases that could degrade DNA during extraction.
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Explain 'sticky ends' in DNA ligation.
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Sticky ends are short single-stranded overhangs produced by restriction enzyme cuts, allowing DNA fragments to anneal together easily.
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What is genomic DNA?
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Genomic DNA is the complete DNA content of an organism, containing both coding (genes) and non-coding regions.
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What is the significance of gene cloning in agriculture?
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Gene cloning is used to create genetically modified crops that can resist pests, diseases, and environmental stresses.
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Describe the term 'nucleic acids'.
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Nucleic acids are biopolymers (DNA and RNA) essential for the storage, transmission, and expression of genetic information.
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What role does sodium dodecyl sulfate (SDS) play?
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SDS is a detergent used to lyse cell membranes and denature proteins during nucleic acid extraction.
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Differentiate between exonuclease and endonuclease.
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Exonuclease removes nucleotides from the ends of DNA, while endonuclease cuts internal phosphodiester bonds within a DNA strand.
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What is a genome library?
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A genome library is a collection of cloned DNA fragments from a particular organism, used for gene identification and research.