Cloning and analysis of immunoglobulin light chain genes
Replenishment date: 30.01.2009
Content: 90130150015537.zip (45.28 KB)
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Description
Cloning and analysis of sterlet immunoglobulin light chain genes
TABLE OF CONTENTS
LIST OF ABBREVIATIONS .............. 3
INTRODUCTION .................................. 4
LITERATURE REVIEW ................. 5
STRUCTURE OF IMMUNOGLOBULINS ...................................... 5
GENOMIC ORGANIZATION AND EXPRESSION
GENES OF IG L-CHAINS IN MAMMALS ........................... 7
GENOMIC ORGANIZATION AND EXPRESSION
GENES OF IG L-CHAINS IN LOWER VERTEBRATES ................. 11
EVOLUTION OF IH L-CHAIN GENES ........ 19
MATERIALS AND METHODS .......... 24
DNA ELECTROPHORESIS ................ 24
ISOLATION OF DNA FROM GEL ............. 24
RECEIVING RADIOACTIVE PROBES ............................ 25
ISOLATION OF PLASMID DNA ................................. 26
PURIFICATION OF PLASMID DNA BY THE EQUILIBRIUM METHOD
CENTRIFUGATION IN THE CsCl DENSITY GRADIENT ......... 26
TRANSFORMATION OF PROKARYOTIC CELLS ............ 27
ISOLATION OF LEUKOCYTES FROM FISH BLOOD .......................... 28
ISOLATION OF TOTAL RNA FROM LEUKOCYTES ............ 28
ISOLATION OF HIGH-MOLECULAR DNA
FROM EUKARYOTIC CELLS ............. 28
CONSTRUCTION OF A cDNA LIBRARY ........................... 29
POLYMERASE CHAIN REACTION ................................... 33
DNA SUBCLONING ........................ 34
CDNA LIBRARY SCREENING ..................................... 34
DETERMINATION OF DNA NUCLEOTIDE SEQUENCE ................... 35
SAUZERN BLOT HYBRIDIZATION ..................... 36
RESULTS ................................... 38
CONSTRUCTION OF A cDNA LIBRARY AND ITS SCREENING ................. 38
DEFINITION OF THE PRIMARY STRUCTURE .......................... 40
COMPARATIVE ANALYSIS OF THE PRIMARY STRUCTURE ............................. 40
ANALYSIS OF THE GENOMIC ORGANIZATION .......................................... 41
DISCUSSION ............................. 46
CONCLUSIONS .......................... 48
REFERENCES ......................... 49
TABLE OF CONTENTS
LIST OF ABBREVIATIONS .............. 3
INTRODUCTION .................................. 4
LITERATURE REVIEW ................. 5
STRUCTURE OF IMMUNOGLOBULINS ...................................... 5
GENOMIC ORGANIZATION AND EXPRESSION
GENES OF IG L-CHAINS IN MAMMALS ........................... 7
GENOMIC ORGANIZATION AND EXPRESSION
GENES OF IG L-CHAINS IN LOWER VERTEBRATES ................. 11
EVOLUTION OF IH L-CHAIN GENES ........ 19
MATERIALS AND METHODS .......... 24
DNA ELECTROPHORESIS ................ 24
ISOLATION OF DNA FROM GEL ............. 24
RECEIVING RADIOACTIVE PROBES ............................ 25
ISOLATION OF PLASMID DNA ................................. 26
PURIFICATION OF PLASMID DNA BY THE EQUILIBRIUM METHOD
CENTRIFUGATION IN THE CsCl DENSITY GRADIENT ......... 26
TRANSFORMATION OF PROKARYOTIC CELLS ............ 27
ISOLATION OF LEUKOCYTES FROM FISH BLOOD .......................... 28
ISOLATION OF TOTAL RNA FROM LEUKOCYTES ............ 28
ISOLATION OF HIGH-MOLECULAR DNA
FROM EUKARYOTIC CELLS ............. 28
CONSTRUCTION OF A cDNA LIBRARY ........................... 29
POLYMERASE CHAIN REACTION ................................... 33
DNA SUBCLONING ........................ 34
CDNA LIBRARY SCREENING ..................................... 34
DETERMINATION OF DNA NUCLEOTIDE SEQUENCE ................... 35
SAUZERN BLOT HYBRIDIZATION ..................... 36
RESULTS ................................... 38
CONSTRUCTION OF A cDNA LIBRARY AND ITS SCREENING ................. 38
DEFINITION OF THE PRIMARY STRUCTURE .......................... 40
COMPARATIVE ANALYSIS OF THE PRIMARY STRUCTURE ............................. 40
ANALYSIS OF THE GENOMIC ORGANIZATION .......................................... 41
DISCUSSION ............................. 46
CONCLUSIONS .......................... 48
REFERENCES ......................... 49