PUKYONG

GUIDE OAK 국가리포지터리 부경대학교

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Abstract: Microalgae show the ability to convert atmospheric CO2 into more complex and commercially valuable organic compounds. Many species and strains of spirulina (A. platensis) have been isolated worldwide and made useful in a variety of fundamental and applied research studies: commercial mass cultures have indeed been developed for the food industry in local areas but also for alternative biofuel feedstock, skin-care product resources. Thus, the goal of this study was to isolate mutant strains of spirulina (A. platensis) by chemical mutation in order to produce anti-oxidative compounds. And then, the mutants were checked by SOD to select those with higher SOD activity than the wild type. Furthermore, the mutation was checked at molecular level by PCR cloning, sequencing in order to find the mutated gene or genes. As results, two mutants were identified to have higher SOD activity than the wild type. The mutants NMU-1-9 and NMU-2-5 showed 4.20% and 27.31% respectively increase in SOD activity compared with the wild type. However, in comparison with the wild type, those mutants didn't show any significant difference in growth rate, chlorophyll and carotenoid contents. The partial SOD gene was found and was identical with the one in the gene bank. However, the molecular characteristics of the full sequence of the SOD gene are unknown so far.

URI: https://repository.pknu.ac.kr:8443/handle/2021.oak/1379
http://pknu.dcollection.net/jsp/common/DcLoOrgPer.jsp?sItemId=000001966788

Table Of Contents: List of figures iv
List of tables v
Abstract vi
I. Introduction 1
II. Materials and methods 5
2.1 Spirulina culture and growth conditions 5
2.2. Isolation of mutant strains 8
2.2.1. Mutagenesis and selection of mutants 8
2.2.2. Biomass analysis 9
2.2.3. Pigment extraction and quantification 10
2.2.4. Antioxidant enzyme activity 11
2.3. Molecular characterization of Arthrospira platensis 12
2.3.1. Isolation of genomic DNA 12
2.3.2. Purification of genomic DNA 14
2.3.3. DNA quantification 15
2.3.4. DNA quality analysis 16
2.3.5. PCR amplification and identification of SOD gene 16
2.3.6. Construction of a spirulina fosmid library 20
2.3.7. Screening of the fosmid library 20
III. Results 22
3.1. Isolation of A. platensis mutants 22
3.2. Growth of wilt type and mutants of A. platensis 24
3.3. Colony morphology of mutant strains of A. platensis 26
3.4. Chlorophyll content of wild type and mutants of A. platensis 28
3.5. Carotenoid content of wild type and mutants A. platensis 30
3.6. SOD activity of wild type and mutants A. platensis 32
3.7. DNA quality, quantity and purity 34
3.8. PCR amplification 36
3.9. Selection of positive clone and Sequencing 38
IV. Discussion 40
V. Conclusion 43
VI. Acknowledgements 44
VII. References 45

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