The expression of plant vicilin DNA in yeast

Abstract

The aim of this project was to study the expression of recombinant DNA plasmids containing the plant seed protein gene for vlcilin, in the yeast saccharomyces cerevisiae. The fidelity of expression was detected by a complete functional protein as judged by binding to anti-vicilin anti-body. This allowed one to gain an insight into the molecular genetics of yeast, especially for expressing heterologous plant proteins. The aim can be further subdivided into a variety of studies; (i) the expression of a genomic vicilin clone, containing the introns and using a yeast promoter, in the yeast, (ii) the expression of a cDNA vicilin clone, which contains no leader sequences, (ill) a study of the transformation systems used for yeast, and (iv) the analysis of the expression using electronmicroscopy, and the study of the secretory pathway involved. The results obtained in this project can be elucidated in the relevant sections of the text. But the production of the genomic clone had to be abandoned as obtaining the genomic DNA proved to be too difficult, as the time allocated to the project was limited. The cDNA vicilin plasmid was constructed, but again due to the limited time, the plasmid could not be transformed into the yeast and analysed. The study of the transformation systems proved to be successful, except the genomic and cDNA clones were not studied. Using the electronmicroscope. The micrographs obtained were only of the yeast containing the plasmid pDUB2018.which contains the same cDNA sequences used in (ii). These showed that the leaderless cDNA was expressed and could be seen to be partially associated with membrane structures.