Biology Chap 11
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| 11714887093 | Biotechnology | The integration of natural science and organisms, cells, parts thereof, and molecular analogues for products and services. | 0 | |
| 11714956511 | Genetic Engineering | Techniques to alter the chemistry of genetic material to introduce these into host organisms and thus change the phenotype of host organisms. | 1 | |
| 11715103985 | 2 Principles of biotech | Genetic engineering Maintenance of sterile ambiance to enable growth of only the desired microbe in large quantities to help in manufacture of biotech products. | 2 | |
| 11715121605 | Characteristic feature of asexual and sexual reproduction | Sexual- permits variation by unique combinations of genetic setup Asexual- Preserve genetic info | 3 | |
| 11715130286 | 2 genetic engineering techniques that involve creation of recombinant DNA. | Gene cloning Gene transfer | 4 | |
| 11715133442 | Why is genetic engineering preferred over traditional hybridisation? | Involves inclusion and multiplication of undesirable genes too Genetic engineering techniques introduce only 1 set of desirable genes in target organism. | 5 | |
| 11715145141 | What prob occurs if an alien DNA enters our body? How can this prob be rectified? | Not be able to multiply itself in progeny cells of organism. Can be rectified if alien DNA is part of a chromosome of host called origin of replication and thus, replicated with it. | 6 | |
| 11715850713 | Cloning | Making multiple identical copies of template DNA | 7 | |
| 11715913091 | First instance of construction of an artificial recombinant DNA molecule | Linking a gene encoding antibiotic resistance with a native plasmid of Salmonella Typhimurium | 8 | |
| 11715917755 | Plasmid | Autonomously replicating circular extra-chromosomal DNA Eg- pBR322 cloning vector | 9 | |
| 11715926496 | Scientists who formed the first recombinant DNA | Stanley Cohen Herbert Boyer 1972 | 10 | |
| 11715938459 | Molecular scissors | Restriction enzymes Cutting of DNA at specific locations | 11 | |
| 11715957324 | Function of DNA ligase in genetic modification | Acts on cut DNA molecules and joins their ends Eg- Linking of antibiotic resistance gene with plasmid vector | 12 | |
| 11715948573 | Function of plasmid DNA | Act as vectors to transfer the piece of DNA attached to it. | 13 | |
| 11715969540 | Recombinant DNA | New combination of circular autonomously replicating DNA created in vitro after genetic modification | 14 | |
| 11715982897 | Basic steps of genetic modification of an organism | Identification of DNA with desirable gene Introduction of identified DNA into host Maintenance of introduced DNA in host Transfer of DNA to its progeny | 15 | |
| 11716012040 | 2 enzymes responsible for restricting growth of Bacteriophages in EColi. | 1963 Milestone of biotechnology Methyl transferase- added methyl grps to DNA to protect it from fragmentation Restriction endonucleases- Fragmentation of DNA | 16 | |
| 11716034133 | 1st restriction endonuclease isolated | 1968 Hind II Cut DNA molecule at particular point by recognizing a specific 6 b.p. sequence. | 17 | |
| 11716044313 | Recognition sequence | Specific base pair sequence at which a restriction enzyme cuts the DNA | 18 | |
| 11716053554 | Naming of restriction enzyme | 1st letter- genus 2nd n 3rd letter- species of prokaryotic cell frm which they were isolated 4th letter- name of strain Roman no. after name- order of isolation of enzymes frm the strain of bacteria. | 19 | |
| 11716070404 | Naming of Hind II | H- haemophilus in- influenzae d- d-type II- 2nd isolated RE | 20 | |
| 11716076443 | Isolation of EcoRI | Escherichia coli RY 13 | 21 | |
| 11716087518 | Exonucleases | Remove nucleotides from the ends of DNA | 22 | |
| 11716088752 | Endonucleases | Make cuts at specific positions within the DNA | 23 | |
| 11716096235 | How does a restriction endonuclease function? | Inspects length of DNA sequence Finds specific recognition sequence Bind to DNA and cut each strand of double helix at specific points | 24 | |
| 11716124204 | Palindromic nucleotide sequences | Palindrome in DNA is a sequence of base pairs that reads the same on 2 strands when orientation of reading is kept the same. Eg- 3'-->5' orientation | 25 | |
| 11716139778 | At what point do REs cut DNA? | Cut the strand of DNA a little away from the centre of palindrome sites , but b/w the same 2 bases on the opposite strands | 26 | |
| 11716153453 | Sticky ends | Overhanging stretches on each strand after action of REs. | 27 | |
| 11716157426 | Why are sticky ends called so? | Form H-bonds with their complementary cut counterparts | 28 | |
| 11716161411 | Use of sticky ends | Facilitates action of enzyme DNA ligase | 29 | |
| 11716177524 | Use of REs | In genetic engineering to form recombinant molecules of DNA. | 30 | |
| 11716191312 | Why should vector and foreign DNA be cut by same RE? | Same kind of sticky ends on both Joined 2gether end to end using DNA ligase. | 31 | |
| 11716211207 | How are DNA fragments separated? | By gel electrophoresis | 32 | |
| 11716213927 | Gel Electrophoresis | Technique used to separate biomolecules under electric field based on their size | 33 | |
| 11716223698 | Principle behind gel electrophoresis | DNA fragments- -vely charged Forcing them to move to anode under electric field thru matrix Smaller fragment, farther it moves | 34 | |
| 11716238429 | Commonly used matrix | Agarose gel Natural polymer extracted frm sea weeds Its sieving effect causes separation of fragments. | 35 | |
| 11716407242 | Stain used in gel electrophoresis | Ethidium bromide | 36 | |
| 11716417101 | How to see DNA fragments in visible light? | Staining of DNA Exposure to UV radiations Bright orange coloured bands seen | 37 | |
| 11716423733 | Elution | Separated bands of DNA are cut out from agarose gel and extracted frm gel piece | 38 | |
| 11716442012 | Why does an alien DNA need a vector? | Multiplication of alien DNA numbers to equal copy no. of plasmid or bacteriophage. Help easy linkage of foreign DNA Easy selection of recombinants frm non- recombinants. | 39 | |
| 11716490608 | Features required to facilitate cloning in a vector | Origin of replication that supports high copy no. Selectable marker Cloning sites | 40 | |
| 11716501820 | Origin of replication | Specific DNA sequence responsible for initiating replication | 41 | |
| 11716511440 | Uses of ori | Piece of DNA linked to this sequence can replicate within host cells Controls copy no. of linked DNA | 42 | |
| 11716537694 | Selectable markers | A gene introduced into a vector, that confers a trait suitable for artificial selection. | 43 | |
| 11716611088 | Use of selectable marker | Identifying and eliminating non-transformants Selectively permitting the growth of transformants | 44 | |
| 11716618114 | Transformation | Process by which piece of DNA is introduced in host bacterium. | 45 | |
| 11716633954 | Why shld there be few or preferably 1 recognition site in vectors? | More than 1 recognition sites in vector will generate several fragments Complicates gene cloning. | 46 | |
| 11716652196 | Where is ligation of alien DNA carried out? | Restriction site present in one of 2 antibiotic resistance genes. | 47 | |
| 11716676922 | Selection of recombinants based on antibiotic resistance | Recombinants selected based on antibiotic resistant nature Non- recombinants- Resistant to ampicillin, tetracyclin Recombinants- Resistant to ampicillin | 48 | |
| 11716704891 | How to isolate recombinants based on antibiotic resistance? | Plating of recombinants and non- recombinants in ampicillin and tetracyclin containing medium. Ampicillin- Both survive Tetracycline- Recombinants can't survive | 49 | |
| 11716799328 | Why is insertional inactivation a better method of selection of recombinants? | Selection by antibiotic resistance is a time-consuming process as it req. simultaneous plating on 2 plates having diff. antibiotics. | 50 | |
| 11716904126 | Insertional inactivation | Easy method of isolation of recombinants by colour production | 51 | |
| 11716973179 | Principle behind insertional inactivation | Usage of selectable markers which can differentiate recombinants and non- recombinats on basis of ability to produce color in the presence of chromogenic substrate. | 52 | |
| 11716922255 | Procedure of insertional inactivation | Insert incorporated in coding sequence of beta- galactosidase gene Recombinants- colorless due to inactivation of beta-galactosidase gene Non- recombinants- produce blue color in presence of chromogenic substance | 53 | |
| 11717013232 | Why is insertional inactivation called so? | Insertion of foreign gene inactivates the enzyme | 54 | |
| 11717042226 | Agrobacterium tumifaciens | Soil bacterium Pathogen of dicot plants Uses T-DNA to transform normal plant cells into tumour | 55 | |
| 11717090749 | Ti plasmid | Tumour inducing plasmid Used if host is a plant cell T-DNA present in this Tumour inducing property is inactivated and it is now used as a cloning vector | 56 | |
| 11717108051 | Ti plasmid is called natural gene engineer | During infection, Ti plasmid induced and tumor formed Due to this specific property of altering gene. | 57 | |
| 11717120252 | Retrovirus | Ability to transform normal cells into cancerous cells Disarmed and used to deliver desirable genes in animal cells. | 58 | |
| 11717326545 | Why can't DNA pass thru cell membranes? | Hydrophilic molecule The bacterial cells must be made competent to take up DNA | 59 | |
| 11717336017 | Heat shock | Treat bacteria with specific conc. of Ca Recombinant DNA forced in cells by putting cells and DNA on ice, 42' conditions and ice again. | 60 | |
| 11717358913 | Why are bacterial cells treated with Ca in heat shock method? | Method of insertion of recombinant DNA in host Increases efficiency with which DNA enters bacteria thru pores in cell wall. | 61 | |
| 11717367010 | Micro injection | Method of insertion of recombinant DNA in host Recombinant DNA directly injected into nucleus of animal cell | 62 | |
| 11717373297 | Biolistics/ Gene Gun | Method of insertion of recombinant DNA in host Cells are bombarded with high velocity micro particles of gold or tungsten coated with DNA Suitable for plants | 63 | |
| 11717383424 | Usage of disarmed pathogen vectors | Method of insertion of recombinant DNA in host Disarmed pathogen vectors infect the cell and transfer recombinant DNA in host | 64 | |
| 11717409747 | Processes of Recombinant DNA Technology | Isolation of DNA Fragmentation of DNA by REs Isolation of desired gene fragment by gel electrophoresis Ligation of DNA fragment in vector Transfer of recombinant DNA in host Culturing host cells in a medium on large scale Extraction of desired product | 65 | |
| 11717439275 | Isolation of bacterial DNA | Lysozyme | 66 | |
| 11717443023 | Isolation of plant DNA | Cellulase | 67 | |
| 11717445868 | Isolation of fungal DNA | Chitinase | 68 | |
| 11717452623 | Removal of RNA | Treatment with ribonuclease | 69 | |
| 11717461304 | Removal of Proteins | Treatment with proteases | 70 | |
| 11717464876 | Removal of lipids | Treatment with lipases | 71 | |
| 11717470444 | How to obtain purified DNA after isolation? | Addition of chilled ethanol leads to precipitation of purified DNA Collection of fine threads in suspension | 72 | |
| 11717486409 | How are restriction enzyme digestions performed? | Incubating purified DNA molecules with restriction enzyme at optimal conditions req. for that specific enzyme Electrophoresis to chk progression of digestion. After cutting by REs, ligation gv rDNA | 73 | |
| 11717532144 | Polymerase chain reaction (PCR) | Multiple copies of gene of interest is synthesised in vitro using 2 sets of primers and DNA polymerase enzyme. | 74 | |
| 11717550243 | Primers | Small chemically synthesised oligonucleotides tht are complementary to regions of DNA. | 75 | |
| 11717561636 | Use of DNA polymerase enzyme | Extends primers by use of nucleotides in reaction and genomic DNA as template. | 76 | |
| 11717568765 | Thermostable DNA polymerase | -Helps in repeated amplification of DNA by repeated replication of DNA -Isolated frm Thermus aquaticus bacterium -Remains active during high temp. induced denaturation step of PCR. | 77 | |
| 11717596015 | How to induce further cloning of recombinant DNA? | Amplified fragment used to ligate with vector. | 78 | |
| 11717611323 | Why is there a need for recombinant DNA to be expressed? | In almost all recombinant technologies, ultimate is to produce proteins, which is not possible unless DNA gets assimilated in host cells | 79 | |
| 11717657651 | Why is there a need for large scale production of recombinant DNA? | Large amount of target proteins needed. | 80 | |
| 11717659855 | Recombinant protein | Protein encoding gene is expressed in a heterologous host. | 81 | |
| 11717674033 | How to produce higher yield of desired protein? | Cont. culture system- Used medium drained frm 1 side and fresh medium enters frm other. Maintains cells in physiological active exponential phase. Produces larger biomass | 82 | |
| 11717688543 | Req. for bioreactors | Small volume cultures cannot yield appreciable quantities of products Large volumes of culture can be processed (100-1000L) | 83 | |
| 11717713253 | Bioreactors | Vessels in which raw materials are biologically converted into specific products, individual enzymes, etc. using microbial plant, animals or human cells | 84 | |
| 11717729081 | Use of bioreactors | Provides optimum growth conditions (temp., pH, substrate, salts, vitamins, O2) to achieve product | 85 | |
| 11717775875 | Stirred- tank reactor | cylindrical or curved base- facilitates mixing of contents Stirrer- facilitates even mixing and O2 availability thru bioreactor Air can be bubbled thru reactor Agitator system O2 delivery system Foam control system Temp. control system pH control system Sampling ports- small vol. of culture withdrawn periodically | 86 | |
| 11717804975 | Downstream processing | Series of separation and purification processes that protein has to go thru b4 marketing Formulated with suitable preservatives. Strict quality control testing | 87 |
