文摘
英文文摘
ACKNOWLEDGEMENTS
GENERAL INTRODUCTION: VECTOR-BORNE DISEASES AND TSETSE FLIES
1. Vector-borne Disease Impact
2. Tsetse-transmitted Diseases
3. Control of Vector-borne Diseases
4. Tsetse Biology and Trypanosome Transmission
5. Alternative Vector Control Techniques
6. Focus of the thesis
CHAPTER ONE: IDENTIFICATION AND CHARACTERIZATION OF GENES THAT ARE DIFFERENTIALLY EXPRESSED IN THEMIDGUT TISSUES OF G. M. MORSITANS IN RESPONSE TO BLOOD FEEDING
1. Introduction
2. Materials and methods
2.1 Insects
2.2 Construction of cDNA libraries representing blood meal induced genes
2.3 Selection of gene fragments induced upon blood feeding
2.4 Construction and screening of midgut cDNA library
2.5 Identification of the 5'-end ofGsp2 cDNA
2.6 Southern blot analysis of Gspl and Gsp2
2.7 Temporal regulation of expression of Gspl, Gsp2, GmCatB,GmZmp and GmZcp mRNA during digestion and Expression in different tissues
2.8 .Trypsin and chymotrypsin enzyme assay and protein determination
2.9 Expression analysis in response to parasite-feeding and parasite injection
3. Results
3.1 Isolation and characterization of cDNAs and their putative products
3.2 Genomic analysis
3.3 Gene expression in response to blood-feeding and in different fly tissues
3.4 Analysis of trypsin and chymotrypsin activities in midgut
3.5 Immune responsive nature of gene expression
4. Discussion
CHAPTER TWO: CLONING AND FUNCTIONAL EXPRESSION OF A FAT BODY-SPECIFIC CHITINASE cDNA FROM THE TSETSE FLIES,G. M. MORSITANS
1. Introduction
1.1 Classification of chitinases modules
1.2 Chitinase gene organization and physiology
1.3 Biochemistry of Chitinases
1.4 Chitinases in biological control
2. Materials and Methods
2.1 Insects
2.2 Isolation of chitinase cDNA clone
2.3SouthernBlotAnalysis
2.4 Northern Blot Analysis
2.5 Analysis ofchitinase activity in fly tissues
2.6 Expression of GChitl in vitro
2.7 Detection of the recombinant GChitl (recGChit1) protein
2.8 Purification of the recGChit1 protein
2.9 Determination of pH profiles and allosamidin inhibition curves of recGChit1
2.10 Antibody Preparation and Immunoblotting
3.Results
3.1 Isolation of G. m. morsitans chitinase cDNA
3.2 Genomic organization of GChitl
3.3 Expression of GChitl mRNA
3.4 Detection of chitinase activity in tsetse tissues
3.5 pH profile and allosamidin sensitivity ofrecGChitl
3.6 Immunoblotting analysis of GChitl protein in tsetse
4. Discussion
REFERENCES
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