The Study Of Construction And Production Analysis Of Transgenic Yeast Strain With ZDS

Carotenoids was a kind of important pigment. In the multitude of natural carotenoids,beta-carotene as the pre-body of vitamin A was paid much attention by clinical medicine circles. Recently,the application and study of beta-carotene were promptly developed in the fields of clinical medicine and industry of foodstuff and cosmetic. The purpose of this experiment is the construction of transgenic yeast strain and the analysis and detection of its production. The vector I used in the experiment is pPIC3.5. I construct a new plasmid pPIC3.5-ZDS, which involves a foreign gene zds. After being linearized by BglII, it was integrated into yeast chromosome with electroporation. After screening, I got positive clones, which can express carotenoid highly. Then I proceeded with a serious molecular detection such as PCR, dot-blot, SDS-Page etc. The result showed that it was the foreign that lead to all these changes such as the color and production. To extract carotenoid from the YPD culture, I used a way known as acid-hot method. The result indicated that the quantity of production was 684μg/g(dried yeast body),β-carotene was 285μg/g(dried yeast body). In order to analysis the component and its percent of the carotenoid production, I took on HPLC analysis. I got a satisfied result. I found the production was composed of three component,one of them wasβ-carotene,and I also got the map of abstract spectrum of them. In HPLC detection,I got every components'general percent and their precise position under stated constitution. At last, I did some industrial experiment to make sure the culture and the conditions for the next study.