Dissertation > Excellent graduate degree dissertation topics show

The Cloning of PsbA Promoter from Chlamydomonas Reinhardtii Chloroplast and Its Activity Detection in E.Coli

Author: ZhangWei
Tutor: SuZhongLiang
School: Qingdao University of Science and Technology
Course: Biological Engineering
Keywords: chlamydomonas chloroplast PsbA promoter engineering green fluorescent protein gene
CLC: Q943.2
Type: Master's thesis
Year: 2011
Downloads: 52
Quote: 0
Read: Download Dissertation


Chlamydomonas is used as a model green alga in many studies. Because it’s genome sequence has been sequenced, and the genetic mechanism is clear; it is a eukaryotic unicellular green algae; it is not only a autotrophic organism but also a heterotrophic organism; it contains a big cup chloroplast that can be easily transformed; it has short growth cycle and high photosynthetic efficiency, it is called“green yeaet”and usually is used as host cells to express foreign gene. The Chlamydomonas is the only organism that has been transformed successfully in nucleus, chloroplast and chondriosome. But the poor foreign gene expression level in chloroplast is a bottleneck that limits its using as bioreactor. The one of main factor affecting the foreign gene expression is promoter. The strong promoter can increase the transcription frequency and increase foreign genes expression level. The Chlamydomonas chloroplast endogenic photosystem II protein D1 promoter is usually considered as a strong promoter. In this research, the promoter above mentioned is cloned, and we hope that the promoter may be promote the foreign genes expesson in Chlamydomonas chloroplast.The contents of this study is as following:Firstly, the chlamydomonas total genome is extracted, and the PsbA primer is designed. With total DNA as templates, the PsbA promoter is cloned using the method of PCR.Secondly, the plasmid p64D-PsbA-aadA is constructed. The PsbA promoter is ligated to the spectinomycin resistance gene (aadA) upstream. The construction is transformed into E.coli DH5α. After screening on ampicillin and spectinomycin plate, the positive clone is obtained. Which proves that the PsbA promoter has the promoter activity.Finally, the plasmid psk-PsbA-gfp is constructed. The green fluorescent protein gene (gfp) is ligated to PsbA promoter downstream. After transforming E.coli DH5αand screening, the positive clone is obtained. Using the fluorescent microscope, the green fluorescence is observed in E.coli. This proves that PsbA promoter can regulate the green fluorescent protein gene expression in E.coli. The results of this research lay the foundation for further chlamydomonas chloroplast engineering study.

Related Dissertations

  1. Service Model Driven Architecture (SMDA) Oriented Modeling Techniques and Tool Implemention,TP311.52
  2. Military exercises in the Corps of Engineers engineering equipment maintenance to protect the allocation of resources,E251.2
  3. Cloning and Expression Analysis of GPx, GST and SAHH Genes in Chlamydomonas Sp. ICE-L from Antarctica,Q943.2
  4. The Problems and Countermeasures in Respect to Agent Construction of the Project: Guangzhou Asian Games Venues,G812.2
  5. Research on Setting Innovation Problems of Mathematics in College Entrance Examination,G633.6
  6. Aesthetic Education in Science and Engineering University Current Situation and Countermeasures,G641
  7. Study on Proportions and Applications of Concrete with Iron Mine Tailings,TU528
  8. A Study of Household Handles Design from the Perspective of Kansei Engineering,TS664.01
  9. Dendrobium candidum chloroplast microsatellite development and application of interspecific,S567.239
  10. Acid solution on the structure and properties of chitosan / hydroxyapatite composite,R318.08
  11. Of the main body of ethical responsibility,B82-057
  12. The Interaction of Fluroxypyr with the Green Alga Chlamydonomas Reinhardtii,S482.4
  13. Study on the Treatment of Coke Wastewater with Iron/Carbon/Fenton-biological Combination Processing,X784
  14. Analysis of Gene Ycze in Bacillus and Biocontrol Efficacy of Genetically Engineered Bacillus Expressed Harpin on Rice Bacterial Leaf Blight,S476.1
  15. Development and Application of Biological Seed Coating Formulation for Soybean,S565.1
  16. Genetic Character and Gene Mapping of an Albino Green-Revertible Mutant in Rice (Oryza Sativa L.),S511
  17. The Research on Harbin Engineering University Subject Team Management and Support Mechanisms,G647
  18. Research on Development Modeling for RIA,TP311.5
  19. Construction of Engineering Strain Saccharomyces Cerevisiae Metabolizing Xylose,Q78
  20. Primary Research on Strring Culture and Storing of Human Dermal Fibroblasts,R329
  21. Study of Essential Safety Improve of Zhaizhen Main Mine Production Systems,TD79

CLC: > Biological Sciences > Botany > Plant Cell Genetics > Plant Genetic Engineering
© 2012 www.DissertationTopic.Net  Mobile