I= VARIABILITY ANALYSIS IN IMMUNOGENIC REGION OF SURFACE GENE OF PAKISTANI ISOLATES OF HEPATITIS B VIRUS
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Title of Thesis
VARIABILITY ANALYSIS IN IMMUNOGENIC REGION OF SURFACE GENE OF PAKISTANI ISOLATES OF HEPATITIS B VIRUS
Author(s)
Naaz Abbas
Institute/University/Department Details
University Of The Punjab/ National Center Of Excellence In Molecular Biology University Of The Punjab
Session
2001
Subject
Molecular Biology
Number of Pages
148
Keywords (Extracted from title, table of contents and abstract of thesis)
hepatitis b, pres gene, s gene, epitopes, hbv sequences, pakistanian hepatitis b virus, pres1 sequences

Abstract

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1085.41 KB
S. No. Chapter Title of the Chapters Page Size (KB)
1 Contents
KB
2 1 Introduction 1
66.02 KB
3 2 Literature survey 10
199.68 KB
  Hepatitis B virus 11
  Historical Background 11
  HBV Genome 12
  PreS1 region 17
  PreS and S gene mutants 21
  Mutation reported in PreS1 region 23
  Epidemiology of infection 25
  Transmission 26
  Target Cell for HBV 27
  Genetic analysis of HBV 31
  Phylogenetic analysis 33
  Treatments 37
4 3 Experimental materials 37
213.16 KB
  Composition of reagents 38
  Methods 43
  Collection of HBsAg positive samples 43
  Plasma isolation 43
  Viral DNA extraction 44
  Proteinase K Digestion Method 44
  Nal Extraction method 45
  Incubation with NaOH 46
  PCR Primer design 46
  Preparation of plasmid DNA AS HBV position control 47
  Preparation of competent cell using CaCl 2 47
  Transformation of competent cells by heat shock method 48
  Preparation of plasmid DNA( Alkaline Lysis ) 49
  Spectrophotometer of plasmid DNA 52
  Quantitiation of DNA through minigel 52
  PCR Amplification 53
  Gel Purification of Amplified HBV DNA 57
  Phenol chloroform method 57
  DNA Purification from gel slice using’s Nuclei 58
  DNA purification from low melting gel using Wizard DNA purification 59
  Cloning Amplified HBV DNA 60
  Restriction Enzyme Digestion 61
  DNA Sequencing 62
  Silver Sequencing 62
  Fluorescent Sequencing 64
  Sequencing Variation Homology Analysis 66
  Novelty search though blast 66
  Phylogenetic analysis 67
5 4 Results 68
426.46 KB
  Screening of patients and sampling 68
  Primer Designing 68
  Viral DNA extraction 75
  Position control HBV DNA used in PCR 75
  PCR amplification 79
  Cloning preS region form Pakistanian HBV isolates 83
  Comparison with other subtypes/ genotypes 101
  Pairwise analysis of variability within the Pakistanian HBV sequences 110
  Novelty search through Blast 110
  Phylogenetic Analysis 113
6 5 Discussion 117
102.88 KB
7 6 References 131
160.55 KB