Introduction to Biotechnology PDF

Introduction to Biotechnology Dr. Ayaz K. Mallick This PowerPoint is to illustrate the lectures and should not be used as a substitute for textbooks. You must read the recommended textbooks for complete understanding and preparation for the...

Introduction to Biotechnology Dr. Ayaz K. Mallick This PowerPoint is to illustrate the lectures and should not be used as a substitute for textbooks. You must read the recommended textbooks for complete understanding and preparation for the exams.  Learning Objectives At the end of the lecture the students should be able to Define biotechnology Define Recombinant DNA technology and explain the steps mentioning the role of restriction endonucleases, vectors and plasmids with examples Describe the steps and clinical importance of PCR and Blotting techniques  What is Biotechnology? Biotechnology is the manipulation of living organisms and organic material to serve human needs Examples: – Yeast in bread making and alcohol production – Use of beneficial bacteria (to get antibiotics like penicillin and vaccines) – Cloning of plants and animals – Stem cell research  Some discoveries Antony Van Leeuwenhoek Discovers bacteria using a simple microscope Gregor Mendel Conducted the first genetics experiments using pea plants in the mid 1800s. Often considered the founder of genetics. Louis Pasteur Created the rabies vaccine Robert Hooke Invented the compound light microscope First to observe cells in cork James Watson & Francis Responsible for the discovery of the double Crick helix structure of DNA using X-ray photographs Meselson and Stahl Gave the model to explain that DNA replication is semi conservative Paul Berg first developed recombinant DNA technology, a method for insertion of genetic material from one organism into another.  Recombinant DNA technology ‘Recombinant DNA technology’ is a technique by which genes/DNA segment of one organism is transferred to another living organism. This manipulation of DNA by artificial means is called recombinant DNA technology. An important application of Recombinant DNA technology is commercial preparations of important biomolecules in high yield. For example, growth hormones, insulin, immunoglobulins, vaccines, clotting factors, etc. Jokarout WL.IT t.I plasmid 4 DNA  Recombinant DNA technology 04 bj I.si a s I (Plasmid) I am I weather.JP s r m gone 0 III WN.TO 31 ID Id 4.14 EI.BY Ddia Some important terms  Restriction endonuclease (also called molecular scissors) DNA205 DNA.is  Enzymes that can cleave (cut) DNA at a specific nucleotide sequence called restriction sites. Plays important role in recombinant DNA technology in isolating genes of interest from parent DNA. DNA I  Examples of restriction endonuclease enzymes are: TWHMI.sn send EcoRI (Source: Escherechia Coli) windows Taq I (Source: Thermus Aquaticus) Wbi  Vectors was'wwjÉ  They are carriers uses to introduce a gene of interest from human DNA into bacteria. Examples: plasmids, bacteriophages or cosmids.  Plasmids  Small, circular, extrachromosomal DNA molecules found in bacteria.  It helps in antibiotic resistance to bacteria  Used in Recombinant DNA Technology as it can replicate inside the bacteria independent of the bacterial DNA.  Polymerase Chain Reaction 6 (PCR) Technique for generating large quantities of specified DNA DWAIEHIH wiw.si in vetro it It is a cell-free technique www.TE.ed imma Steps of PCR Step I: Denaturation 535 Step 2: Renaturation 5061 Step 3: Extension or Synthesis  IT I we IT WAY 155,1 DNA Taq Polymerase in thermostable at 95 degree Celsius  Uses of PCR e_merashainreatio sodMI.w Prenatal diagnosis of inherited disease – Sickle cell anemia, thalassemia Diagnosis of viral infections – HIV, Covid-19 Diagnosis of bacterial infections – Tuberculosis t.sD.al Diagnosis of cancer – Cancer of cervix  test tube A oldDNA now Blotting technique co is b f in asseiwt.mt.aoimshltasiisIDNA D.J Alibis jtion Techniques used to immobilize and identify DNA, RNA or proteins Types of blotting – Southern blotting: Identification of DNA – Northern Blotting: Identification of RNA MRNA – Western Blotting: Identification of Proteins It 4 is III at nitrocellulose wie DNA 3 I a 131 4J 24 Idly 5 a my  IIIIIII It waving taws 11816 www.ni 2a okbatW  1997 Dolly, a sheep was the first animal cloned mm from diploid cells ; was produced in Scotland