碩士 / 輔英科技大學 / 生物科技系碩士班 / 99 / The objective of this study is to generate transgenic microalgae capable of producing phytase. Two acid phosphatase gene (PH01) expression vectors, pC35S-PH01driven by the 35S promoter and pHR-PH01 driven by Hsp70A-RbcS2 fusion promoters, were constructed and introduced into Chlorella sp. T-89 by electroporation. Stable transgenic lines were obtained by hygromycin selection. Successful transduction and integration of the transgene into host genomes were detected by chromosomal PCR. Western analysis of the crude extract from several transgenic lines showed that the recombinant acid phosphatase gene was expressed in transgenic Chlorella sp. T-89. In addition, both acid phosphatase activity and phytase activity levels were significantly increased in the transgenic lines S1, S5, H1, H2, compared with that in parental hosts. The results demonstrated that transgenically expressed acid phosphatase from Pichia pastori was funtional in transgenic lines of Chlorella sp. T-89. The heat inducibility of Hsp70A-RbcS2 fusion promoters was confirmed by 2 and 4.6 fold induction of acid phosphatase and phytase activity, respectively, when incubation temperature was increased to 42oC. Taken together, acid phosphatase genes from Pichia pastori were successfully expressed in Chlorella sp. T-89 under the control of a constitutive 35S promoter or heat-inducible Hsp70A-RbcS2 fusion promoter.
| Identifer | oai:union.ndltd.org:TW/099FY005111002 |
| Date | January 2011 |
| Creators | Shih-Yuan Wang, 王識淵 |
| Contributors | Te-Jin Chow, 周德珍 |
| Source Sets | National Digital Library of Theses and Dissertations in Taiwan |
| Language | zh-TW |
| Detected Language | English |
| Type | 學位論文 ; thesis |
| Format | 58 |
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