Date of Award

Spring 1984

Document Type

Thesis

Department

Life & Environmental Sciences

First Advisor

John Christenson

Second Advisor

James Manion

Third Advisor

Henry Burgess

Abstract

Comparative studies of protein expression and DNA similarity were carried out on two recombinant plasmids of Aleutian Disease Virus. ADV-G genome molecularly cloned into plasmid pUC8 is designated pBMl and a Danish isolate of ADV(DK-ADV) also cloned into pUC8 is designated pBM2. The recombinant plasmid pBMl induced expression of three previously recognized and one unrecognized polypeptides in E. coli strain JM103 that were specifically detected by sera of ADV infected mink. The proteins with approximate molecular weights of 2?K, 36K, 49K, and 58K daltons were detected by immune blotting of JMlO3(pBMl). The same set of proteins was detected in JMlO3(pEM2). No specific proteins were seen in JM103 or JMlO3(pUC8) reacted with Mink AD Sera(MAD) or in JMlO3(pBMl) and JMlO3(pEM2) reacted with Normal Mink Sera(NMS) Comparative study of the genomes of the isolates by restriction enzyme analysis, however, revealed a dissimilarity. Both inserts consisted of an EcoRI-Hindlll fragment replacing a smaller segment of the pUC8 plasmid with complementary ends. Similarity of protein expression suggests similar genomes, but digestion with various restriction endonucleases followed by resolution on agarose gel electrophoresis showed dissimilarities between the DNA with the pBM2 insert containing one more restriction site than the pBMl. These studies seemed to indicate the two isolates were genetically different forms of the Aleutian Disease Virus yet code for the same antigenic determinants.

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