Gene cloning
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Gene cloning involves isolating a specific gene from an organism's DNA and copying it. This is done by cutting the DNA into fragments using restriction enzymes, inserting the gene fragments into bacterial plasmids, and transforming the plasmids into bacteria. The transformed bacteria are cultured to form a DNA library containing many copies of the cloned gene. The library is screened to identify the bacteria containing the gene of interest. cDNA libraries are also created using reverse transcriptase to copy mRNA into DNA that can be expressed in bacteria without introns. Gene cloning is important for research but raises some ethical issues regarding antibiotic resistance and ownership of genetic information.
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This document provides an overview of genetic engineering and its applications to microorganisms. It defines genetic engineering as the direct manipulation of an organism's genome using biotechnology. The key steps involved are isolating the gene of interest, inserting it into a vector, introducing the vector into a host cell, and harvesting the gene product from the clone. Common hosts used are bacteria, yeast, plant and animal cells. The document also discusses some tools used in genetic engineering like restriction enzymes and DNA ligase. It outlines several applications of genetic engineering in medicine, research, agriculture and industry. It concludes by noting some ethical and safety concerns regarding genetically modified organisms.
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Bacteria are useful hosts for genetic engineering and recombinant DNA technology because they are easy and cheap to grow, grow quickly, and can be easily manipulated in the laboratory. Their natural plasmids and viruses can act as vectors to insert recombinant DNA. Plasmids are small, circular pieces of DNA that are not essential for bacterial survival but can contain genes that confer traits like antibiotic resistance and mechanisms for genetic transfer between bacteria. Molecular biologists have engineered plasmids with restriction sites to allow insertion of foreign DNA fragments and propagation of those genes within bacteria.
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3. Recombinant DNA is made by cutting DNA with restriction enzymes and ligating pieces into plasmids, which are then inserted into bacteria.
Gene cloning
Gene cloning involves making many identical copies of a gene. It differs from cloning an entire organism. Gene cloning is done to isolate and study particular genes, identify control sequences and mutations, and engineer organisms for specific purposes like insulin production. It is accomplished by extracting DNA from an organism, cutting it into pieces with restriction enzymes, and inserting the gene of interest into bacterial plasmids which are then taken up by bacteria. This allows the gene to be amplified as the bacteria containing the recombinant plasmid multiply. PCR is another method that amplifies DNA fragments in vitro using heat-stable DNA polymerase and repeated heating and cooling cycles.
Recombination Technology
Recombinant DNA technology allows DNA from different species to be isolated, cut, spliced together, and replicated. This creates new "recombinant" DNA molecules. Key steps include using restriction enzymes to cut DNA into fragments, inserting fragments into cloning vectors like plasmids, and transforming host cells to replicate the recombinant DNA. PCR is also used to amplify specific DNA sequences. Recombinant DNA technology has many applications, including producing human proteins, diagnosing genetic diseases, and detecting bacteria and viruses.
Cloning
Genetic engineering involves transferring genes between organisms using recombinant DNA techniques. This allows genes to be isolated, cloned, and moved within and between different species. Cloning a gene involves using restriction enzymes to cut DNA at specific sequences, and DNA ligase to join DNA fragments together. Cloned genes have many research uses such as determining gene sequences, altering phenotypes, and obtaining protein products of genes.
Cloning
Genetic engineering involves transferring genes between organisms using recombinant DNA techniques. This allows genes to be isolated, cloned, and moved within and between different species. Cloning a gene involves using restriction enzymes to cut DNA at specific sequences, and DNA ligase to join DNA fragments together. Cloned genes have many research uses such as determining gene sequences, altering phenotypes, and obtaining protein products of genes.
Bacterial transformation
Bacterial transformation involves bacteria taking up foreign DNA from their environment. This process allows genes to be transferred between bacteria. Transformation is used in DNA cloning to introduce DNA like plasmids containing genes of interest into bacteria. The bacteria are then placed on antibiotic plates to select those containing the plasmid, which allows the gene of interest to be replicated in large quantities for study or use.
Bacterial transformation
Bacterial transformation involves bacteria taking up foreign DNA from their environment. This process allows bacteria to horizontally transfer genes. Transformation is used in DNA cloning to introduce plasmids containing genes of interest into bacteria. The bacteria are then selected using antibiotics to identify those containing the plasmid, which can then be used to produce large quantities of the gene or its protein product.
Recombinant dna technology
Recombinant DNA technology involves transferring genes between organisms using artificial means. It works by combining DNA from different sources into a single molecule. The process involves generating DNA fragments, inserting the fragments into vectors, introducing the vectors into host cells, and selecting clones containing the recombinant DNA. Common tools used include restriction enzymes to cut DNA, vectors like plasmids to carry DNA, bacterial hosts like E. coli, and techniques like transformation and selection to introduce and identify recombinant DNA. Applications include analyzing gene structure, producing pharmaceuticals, genetically modified organisms, and gene therapy.
Recombinant DNA Technology
Recombinant DNA technology allows DNA from different species to be isolated, cut, and spliced together to form new recombinant molecules. Key tools for recombinant DNA technology include restriction enzymes, ligases, polymerases, vectors, and host cells. Recombinant DNA technology has many applications, including producing human insulin and other proteins for medical use, genetically engineering plants for crop improvement, and DNA fingerprinting for criminal investigations.
gene cloning principles an technique
Gene cloning techniques allow scientists to make multiple copies of gene-sized DNA fragments. The basic cloning process involves inserting a foreign gene into a bacterial plasmid, introducing the recombinant plasmid into bacterial cells, and allowing the bacteria to replicate and produce many copies of the gene. Restriction enzymes cut DNA at specific recognition sites, creating sticky ends that allow insertion of a foreign DNA fragment into a plasmid. Recombinant plasmids are then introduced into bacteria by transformation, allowing clones containing the gene of interest to be identified and isolated.
Lecture 20 biotechnology
The document provides an overview of DNA technology and biotechnology. It discusses how DNA cloning allows scientists to make multiple copies of genes and study their structure, expression, and function. Key techniques described include recombinant DNA, restriction enzymes, gel electrophoresis, and DNA sequencing. Applications mentioned include genetic engineering of plants, microorganisms, and animals for research, agriculture, medicine, forensics, and environmental cleanup.
Genetic engineering
This document summarizes recombinant DNA (rDNA) technology and genetic engineering. It discusses how restriction enzymes and DNA ligases are used to cut and paste DNA from different sources to create recombinant DNA molecules. It also describes genetically modified organisms (GMOs) and provides examples of transgenic organisms. Vectors such as plasmids are used to carry and replicate foreign DNA fragments in bacteria. Selection techniques like antibiotic resistance allow identification of transformed bacteria.
R dna ppt
Recombinant DNA (rDNA) technology involves combining DNA molecules from different sources into new combinations. DNA from one organism is cut and joined with DNA from another organism, and the resulting recombinant DNA is inserted into a host cell. This allows genes and DNA fragments to be expressed, amplified, and mass produced. The basic steps are isolation of genetic material, restriction enzyme digestion, amplification via PCR, ligation, insertion into a host cell, and isolation of recombinant cells. rDNA technology has many applications including basic research, production of therapeutic proteins, agriculture, medicine, and industry. Hybridoma technology uses cell fusion to produce monoclonal antibodies that are identical clones of a single parent cell.
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Gene cloning
- 1. Gene Cloning & Creating DNA Libraries
- 5. Whole organisms are cloned too, but differently
- 8. The action of a restriction enzyme , EcoR I Note: EcoR I gives a ‘sticky’ end
- 19. A simplified diagram of transcription in eukaryotes hnRNA = ‘heterogenous nuclear’RNA in the process of being cut and spliced into messenger RNA
- 24. These points and others require research and informed debate- What are Your thoughts? What legislation has been passed here in New Zealand? Who controls work in genetic manipulation?
- 25. References Kreuzer, H., Massey, A.,2001, Recombinant DNA & Biotechnology , ASM Press, Washington Turner, P.C., et al, 1997, Instant Notes n MolecularBiology , Bios, Oxford www.agbiosafety.unl.edu/education/clone.htm http://avery.rutgers.edu/WSSP/Student Scholars/Session12/Session12html http://www.pssc.ttu.edu/pss3421/gene%20cloning%20Strategies.htm http://www.uic.edu/classes/phar/phar331/lecture7 http://www.biology.arizona.edu
- 26. Picture Credits Cc the cat, slide 5 courtesy of Texas A & M University, College of Veterinary Medicine Photos on slide 5, 7,12 & 17, L. Macdonald 2003 Graphics on slides 4, 8 &19, V. Ward, University of Auckland DNA Hybridization, slide 15, courtesy of Texas Tech University DNA cloning & screening, slides 10 & 14, courtesy of The University of Arizona
- 27. Acknowledgements Thanks go to Craig Millar and Andrew Dodd, School of Biological Science, University of Auckland Compiled by Linda Macdonald For NCEA Biology A.S. 3.6 While on a New Zealand Royal Society Science, Mathematics & Technology Teacher Fellowship