Cloning of Schistosoma mansoni adult worm antigens for the prepatent diagnosis of schistosomiasis /
The objective of this work was to identify recombinant proteins reactive with prepatent mouse serum sequence recombinant clones, characterize and test the recombinant protein in immunoassays for the detection of schistosomiasis. Seven clones were purified from the library after immunoscreening with...
| Main Author: | |
|---|---|
| Format: | Thesis Book |
| Language: | English |
| Published: |
[Place of publication not identified] :
[publisher not identified] ;
1995.
|
| Subjects: | |
| Online Access: | http://proxy.library.tamu.edu/login?url=http://proquest.umi.com/pqdweb?did=742535151&sid=1&Fmt=2&clientId=2945&RQT=309&VName=PQD |
| Summary: | The objective of this work was to identify recombinant proteins reactive with prepatent mouse serum sequence recombinant clones, characterize and test the recombinant protein in immunoassays for the detection of schistosomiasis. Seven clones were purified from the library after immunoscreening with prepatent mouse serum. Three clones were identified as actin, calcium binding protein, and cytochrome oxidase subunit II and four remain unidentified. The partial clone SmCBP2, a calcium binding protein was found to be [] into a 0.5 kb mRNA, and Southern blot analysis indicated the gene to be in a single genomic copy, and possibly without intervening sequences. Two actin cDNA actin clones, SmAct and SmAct2, and a genomic actin clone EMBLSmAct5-1 were identified. The genomic clone contained in the 5' untranslated region, consensus sequences CArG and E boxes, possibly involved in tissue and developmental regulation of actin gene expression. Primer extension analysis revealed a possible [] start site of SmAct located at base 52. The [] of SmAct and SmAct2 were found to be of 1.4 kb and 1.75 kb respectively. Southern blot [] indicated actin to be a multi gene [], and both isoforms contained multiple introns. SmCOX2 was found to encode a schistosome mitochondrial cytochrome oxidase subunit II and was [] into a 1.5 kb RNA. Southern blot analysis revealed the gene introns, and to be present at a single copy per genome. RNase protection assay and Northern blot revealed that genes encoding for SmCBP2, SmAct, SmAct2, SmCOX2 and 3SAP2 were differently expressed in cercariae and adult worms. Clone SmCBP2 was expressed in E. coli, and in ELISA it was reactive with infected mouse serum, and prepatent mouse serum when compared to normal mouse serum. Infected and prepatent human sera was more reactive than normal human sera. ELISA results did not correlate with the number of eggs excreted by human patients, and showed some cross-reactivity. In conclusion actin, a calcium binding protein, and cytochrome oxidase subunit II were purified with prepatent mouse serum. These clones were differently expressed in cercariae and adult worms. SmCBP2 was found to be reactive with prepatent mouse and human sera. |
|---|---|
| Item Description: | Vita. "Major Subject: Microbiology". |
| Physical Description: | xii, 218 leaves : illustrations ; 28 cm. Issued also on microfiche from University Microfilms Inc. |
| Bibliography: | Includes bibliographical references. |