Document Type

Thesis - Open Access

Award Date

1978

Degree Name

Master of Science (MS)

Department

Chemistry

First Advisor

D.G. Kenefick

Abstract

Specific rabbit antisera against purified seedling RNase I from both a hardy (Dicktoo) and less-hardy (Tennessee Winter) cultivar of winter barley was obtained using a 1 mg injection schedule. Both antisera formed a single precipitin band on double immunodiffusion and immunoelectrophoresis when reacted with the homologous crude tissue extract. RNase antigen from either cultivar was highly cross-reactive with both antisera. Passive hemagglutination inhibition was used in an attempt to distinguish purified RNase from both cultivars. A consistent difference in anti-RNase serum specificity between cultivars was shown, but the difference observed by this method was not sufficient to conclude structural differences between the two RNase antigens. Immunodiffusion and rocket immunoelectrophoresis were used to qualitatively and quantitatively test the cross-reactivity of protein preparations from various members of the species Hordeum as well as unrelated species of grasses. Both antisera preparations were sufficiently specific for barley RNase that soluble protein preparations from species other than Hordeum showed no cross-reactivity with either antiserum. Only certain species of Hordeum were shown to be cross-reactive. A quantitative method for the determination of RNase in unpurified tissue extracts was developed using a modification of rocket immunoelectrophoresis. The technique modifications include a template-reservoir which allowed detection of 250 ng of RNase in sample volumes up to 50 ul. The quantity of RNase in unpurified protein extracts from a hardy and a less-hardy cultivar of barley was shown to be the same even though the RNase activity has been shown to differ greatly between the two cultivars.

Library of Congress Subject Headings

Barley
Immunospecificity
Serology
Immune serums
Enzymes
Ribosomes

Format

application/pdf

Publisher

South Dakota State University

Included in

Chemistry Commons

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