Cloning and partial sequencing of an actin gene from Alnus Glutinosa

Author

Ledare F. Habera

Date of Award

12-1987

Degree Type

Thesis

Degree Name

Master of Science

Major

Life Sciences

Major Professor

Beth C. Mullin

Committee Members

Gary Stacey, Stuart Riggsby

Abstract

An alder (Alnus glutinosa) genomic library was constructed in bacteriophage lambda and screened with an actin gene probe from the yeast Candida albicans. Four clones were isolated, three of which contained different actin genes. One clone was chosen for further study, and the region of the alder insert containing the hybridizing region was identified by restriction mapping. Two Hind III fragments (1.1 kb and 1.2 kb) were found to contain the actin hybridizing region. The two Hind III fragments have been subcloned into M13 for sequence analysis. Preliminary sequencing analysis has confirmed that the two Hind III fragments do contain portions of an actin gene. Also the deduced amino acid sequence from these fragments has been compared to other plant actin amino acid sequences and extensive amino acid conservation is observed.

Southern hybridization of the yeast probe to genomic blots of alder DNA digested with Eco HI revealed approximately ten hybridizing restriction fragments. Four of these ten bands were accounted for by the four clones from the alder genomic library. The other hybridizng fragments may represent up to six additional actin genes not yet isolated. These results demonstrate that an actin multigene family is present in the alder genome. Actin multigene families have been found in all plant species examined, and the size of the multigene family in alder is similar to that found in other plant species, such as soybean and tomato.

Recommended Citation

Habera, Ledare F., "Cloning and partial sequencing of an actin gene from Alnus Glutinosa. " Master's Thesis, University of Tennessee, 1987.
https://trace.tennessee.edu/utk_gradthes/13485