Use this URL to cite or link to this record in EThOS: http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.649472
Title: Cloning of genes expressed in the salivary glands during the feeding of the tick Amblyomma variegatum
Author: Delroux, K.
Awarding Body: University of Edinburgh
Current Institution: University of Edinburgh
Date of Award: 2005
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Abstract:
The purpose of this study is to characterise molecules that are secreted in tick salivary glands. A cDNA library of salivary glands was constructed in a lambda phage. Immunoscreening was carried out with a) a hyperimmune antiserum obtained from rabbit immunised with salivary gland extract of unfed A variegatum, b) an antiserum from a rabbit exposed to feeding A variegatum; c) an antiserum from calf repeatedly exposed to feeding A variegatum. All antisera recognised proteins of salivary glands extracts on Western blot analysis, but none detected recombinant salivary gland proteins from the cDNA library. A random selection of cDNA clones were sequenced and compared with those from the Amblyomma variegatum Gene Index (AvGI) and the GenBank of non-redundant sequences database (NCBI). Several clones displayed sequence identity with already known sequences from other tick species. One clone displayed 31% identity with a consensus sequence of the AvGI EST, which shared 28% identity with a 36kDa immunosuppressant molecule isolated from Dermacentor andersoni. Screening of the cDNA library, using the Signal Sequence Trap technique, identified a clone with 99% identity with a glycine rich protein from the AvGI database. This gene was designated FLAv41E and had a molecular weight of 30.9kDa. Reverse transcription PCR (polymerase chain reaction) amplification of FLAv41E gene revealed that it is expressed in the salivary glands of A variegatum female from day 3 to 10 of feeding. Its secondary structure revealed domain similarities with a 29kDa cement protein characterised in Haemaphysalis longicornis ticks. The recombinant FLAv41E protein was expressed in insect cells using a baculovirus system. FLAv41E appears to be a cement protein.
Supervisor: Not available Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID: uk.bl.ethos.649472  DOI: Not available
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