Molecular and immunological characterisation of Acanthocheilonema viteae chitinase
نویسنده
چکیده
Table of Contents 1 Introduction 1 1.1 Structure and function of chitinases 1 1.2 Filarial chitinases are potential targets for interventions 4 1.3 A. viteae as a model to investigate parasitological parameters of filarial diseases 6 1.4 Aims and objectives of the study 7 2 Results 9 2.1 Characterisation of A. viteae chitinase genes 9 2.1.1 Determination of the gene structure of A. viteae chitinase genes 9 2.2 Comparative analysis of A. viteae genomic chitinase sequences with genomic data bases of nematodes 18 2.3 A. viteae chitinase transcripts 21 2.3.1 Screening of A. viteae L3 and adult female cDNA libraries for chitinase transcripts 21 2.3.2 Identification of A. viteae chitinase transcripts 23 2.4 Identification of homologous chitinase sequences from nematode databases 25 2.5 Cloning, expression and purification of recombinant A. viteae chitinase 32 2.5.1 Cloning, periplasmatic/ cytoplasmatic expression and purification of N-terminal portion of A. viteae chitinase in the expression vector pET 22 b (+) in E. coli 32 2.5.2 Activity of N-terminal chitinases 34 2.6 Immunisation studies with A. viteae chitinase in Meriones unguiculatus 35 2.6.1 Immunisation studies with A. viteae N-terminal chitinase fragment 36 2.6.2 Immunisation studies with synthetic peptides from the active site of A.
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