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SubCode & Name : BT9222 AdvancedGenetic Engineering Unit: I Branch: Biotechnology Semester: II Semester
UNITICLONINGANDEXPRESSIONOFGENES: 10
Cloning vehicles, restriction enzymes, restriction modification, linkers, adaptors, homopolymeric tailing,restriction mapping. Expression and purification of recombinant proteins, prokaryotic and eukaryotic expression vectors, in vivo homologous recombination, large scale expression and purificationofproteins.
S.No / Topics to be covered / Time(minutes) / Text book and page numbers / Teaching aids1 / Cloning vehicles / 50 / TB: 1 (43-63)
TB : 3 (57-86) / BB & Chalk
2 / Restriction enzymes / 50 / TB : 3 (49-56) / BB & Chalk
3 / Restriction modification, / 50 / TB : 1 (26-38) / Smart Board
4 / Linkers, Adaptors & Homopolymeric tailing / 50 / TB : 1 (38-41) / Smart Board
5 / Restriction Mapping / 50 / TB : 3 (53-55) / BB & Chalk
6 / Expression and purification of recombinant proteins / 50 / TB : 1 (76-83)
RB: 3 (178-195) / BB & Chalk
7 / Prokaryotic expression vectors / 50 / TB : 3 (121-161)
TB: 1 (70-76) / BB & Chalk
8 / Eukaryotic expression vectors / 50 / TB : 3 (163-189) / Smart Board
9 / In vivo homologous recombination / 50 / TB : 1 (155)
TB: 3 (168- 177) / BB & Chalk
10 / Large scale expression and purificationofproteins / TB : 3 (481-506) / BB & Chalk
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SubCode & Name : BT9222 AdvancedGenetic Engineering Unit: II Branch: Biotechnology Semester: II Semester
UNITIILIBRARY CONSTRUCTION: 8
cDNAgenomicDNAlibraryconstructionandscreening,preparationofDNA,RNA probesimmunoscreeningandblottingtechniques,etc
S. No / Topics to be covered / Time in minutes / Text Book and page Number / Teaching aids11 / GenomicDNAlibrary / 50 / TB: 1 (85-91) / SMART BOARD
12 / cDNA library / 50 / TB: 1 (92-100) / BB & CHALK
13 / PreparationofDNARNA Probes / 50 / TB : 1 (71-73)
TB: 3 (67-72) / BB & CHALK
14 / Screening strategies , probe Design / 50 / TB : 1 (102-103) TB : 1 (105-107) / SMART BOARD
15 / Screening by PCR / 50 / TB: 1 (108-109) / BB & CHALK
16 / Screening by Immunological Assay / 50 / TB: 1 (109-111) / BB & CHALK
17 / Screening by Protein Activity and Functional Complementation and Gain of Function / 50 / TB: 1 (110-114) / SMART BOARD
18 / Blottingtechniques / 50 / TB: 1 (11-16) / BB & CHALK
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SubCode & Name : BT9222 AdvancedGenetic Engineering Unit: III Branch: Biotechnology Semester: II Semester
UNITIII SEQUENCING: 10
Methodology – Chemical & Enzymatic, Automated sequence, Genome sequencing methods–Topdownapproach,Bottomupapproach
S. NO / Topics to be covered / Time in minutes / Text Bookand
page number / Teaching Aids
19 / Chemical Sequencing or Maxam and Gilbert Sequencing / 50 / RB: 3 (560-564) / Smart Board
20 / Enzymatic Sequencing or Sangers Dideoxy Sequencing / 50 / TB: 1 (120-122)
TB: 3 (102-105) / BB & Chalk
21 / Automated Sequencing / 50 / TB: 1 (124-125)
TB: 3 (106-107) / BB & Chalk
22 / Pyrosequencing / 50 / RB: 4 (353-354) / Smart Board
23 / MPSS / 50 / RB: 4 (411-412) / BB & Chalk
24 / Radiation Hybrid mapping / 50 / RB: 4 (358- 360) / BB & Chalk
25 / Happy Mapping / 50 / RB: 4 (360-362) / Smart Board
26 / Optical Mapping / 50 / RB: 4 (354-358) / BB & Chalk
27 / Genome sequencing methods–Topdownapproach or Hierarchial sequencing / 50 / RB: 4(362-364) / BB & Chalk
28 / Genome sequencing methods– Bottomupapproach or Whole Genome Shot Gun sequencing / 50 / RB: 4 (364-371) / Smart Board
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SubCode & Name : BT9222 AdvancedGenetic Engineering Unit: IV Branch: Biotechnology Semester:II Semester
UNITIV PCRANDMUTAGENESIS 07
PCR-Principle,Applications,DifferenttypesofPCR,mutagenesisandchimericprotein engineeringbyPCR,RACE,Kunkels’ methodofmutagenesis.
S. No / Topics to be covered / Time in minutes / Text Book and Reference No / Teaching aids29 / PCR- Principle and Applications / 50 / TB: 1 (19-21)
TB : 3 (110-119) / SMART BOARD
30 / Types of PCR – Real time PCR / 50 / TB: 1 (23-25) / BB & Chalk
31 / Long Accurate PCR, Reverse Transcription PCR, / 50 / TB: 1 (21-23) / Smart Board
32 / MutagenesisbyPCR / 50 / TB: 3 (190-205) / BB & Chalk
33 / Chimericprotein engineeringbyPCR – Case studies / 50 / TB: 3 (205-223) / Smart Board
34 / RACE PCR (5’ and 3’) / 50 / TB: 1 (101-102) / BB & Chalk
35 / Kunkels’ methodofmutagenesis. / 50 / TB: 1 (193-195) / BB & CHALK
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SubCode & Name : BT9222 AdvancedGenetic Engineering Unit: V Branch: Biotechnology Semester: II Semester
UNITVGENETRANSFER GENETHERAPY 10
Introductionofforeigngenesintoplantandanimalcells,Creationoftransgenicplants andanimal knockouts,Genetherapy,Typesandvectors
S. NO / Topics to be covered / Time in minutes / Text Book and page numbers / Teaching aids36 / Introductionofforeigngenes into animal cells and plant cells / 50 / TB : 1 (174-202)
TB : 1 (221-236)
RB : 2 (66-75) / Smart Board
37 / Vectors for plant and Animal, Creationoftransgenicplants and Knockout animals / 50 / TB : 1 (236-246)
TB : 1 (202-220)
TB : 3 (538-590)
TB : 3 (594-626)
RB : 2 (79-100)
RB : 2 (103-302) / BB & Chalk
38 / Genetherapy and types / 50 / RB: 1(1-19) / Smart Board
39 / Viral delivery systems for gene therapy / RB: 1(21-38) / Smart Board
40 / Non viral delivery systems for gene therapy / 50 / RB: 1 (43-60) / BB & Chalk
41 / Targeting of Gene Delivery systems / 50 / RB: 1 (125-139) / BB & Chalk
42 / Gene therapy for multifactorial genetic disorders / 50 / RB: 1 (83-98) / Smart Board
43 / Gene Therapy for infectious Diseases / 50 / RB: 1 (99-122) / BB & Chalk
44 / Ethical Issues in Gene Therapy / 50 / RB: 1 (155-162) / BB & Chalk
45 / Prospects for Gene Therapy / 50 / RB: 1 (163-165) / Smart Board
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SubCode & Name : BT9222 AdvancedGenetic Engineering Unit: V Branch: Biotechnology Semester: II Semester
Course Delivery Plan
Week / 1 / 2 / 3 / 4 / 5 / 6 / 7 / 8 / 9 / 10 / 11 / 12 / 13 / 14 / 15Units / I Unit II Unit III Unit IV Unit V Unit
Text books:
- PrimroseS.B..,TwymanR.H.andOldR.W.PrinciplesofGeneManipulation,6thedition , BlackwellScience, 2001
- Winnacker E.L. FromeGenes to clones: Introduction toGene Technology, Panima,2003
- GlickB.R. & PasternakJ.J.MolecularBiotechnology: Principlesand applicationsofrecombinantDNA,3rded.,ASMPress,2003
- Lemonie,N.R.andCooper,D.N.Genetherapy,BIOSScientific,1996
Reference books:
1.Lemonie, N.R. Understanding Gene therapy, BIOS Scientific, 1999.
2.Adrian slater, Nigel Scott, and Mark Fowler. Plant Biotechnology- The genetic manipulation of plants. Oxford 2004.
3.ALLAN M. MAXAM AND WALTER GILBERT, A new method for sequencing DNA,PNAs, Vol. 74, No. 2, pp. 560-564, February 1977
4.PrimroseS.B..,TwymanR.H.PrinciplesofGeneManipulation and Genomics,7thedition , BlackwellScience, 2006
Prepared by / Approved bySignature
Name / J. Hariharan / Prof. Sulochana Somasundaram
Designation / Asst professor / HOD of Biotechnology
Date / 10-02-13 / 10-02-13
DOC/LP/01/28.02.02
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Date: 10/02/13
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SubCode & Name : BT9222 AdvancedGenetic Engineeing Branch: Biotechnology Semester: II
OBJECTIVE OF THE COURSE:
The aim of the course is to understand the advance techniques in recombinant DNA technology mainly on different types of PCR, automated DNA sequencing, production of chimeric proteins, restriction mapping, different types of screening techniques, large scale production and purification of recombinant proteins, PCR based mutagenesis, gene therapy, blotting techniques, gene transfer methods, production of genetically modified plant and animals, and application of bioinformatics in genetic engineering.
Since the students already learnt the basic techniques of genetic engineering namely restriction enzymes, cloning vectors, cDNA library and application of genetic engineering at the Undergraduate level, the advanced genetic engineering covering all the above mentioned topics would be very useful for Post graduate students to update their knowledge for the research programme.