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Isfahan University of Medical Sciences
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| Document Type
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Latin Dissertation
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| Language of Document
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English
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| Record Number
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102614
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| Doc. No
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T10089
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| Call number
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QW,131,J25c,2006
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| Main Entry
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Jahanian Najafabadi, Ali
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| Title & Author
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Cloning of Polyhydroxyalkanoate synthase genes of pseudomonas aeroginusa PTCC 1310
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| College
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Schools, Pharmacy
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| Date
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, 2006
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| Degree
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Biotechnology, MD
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| Page No
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VI, 73, xx p.: ill ( som col ), Tab
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| Note
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This thesis is also a research project with project ID: 384169
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| Abstract
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Introduction: Over the years, the use of plastics has complicated the problem of disposal of solid wastes. One of the strategies to reduce plastic waste is the use of biodegradable plastics. A group of these plastics are polyhydroxyalkanoates (PHA). To date more than 250 different microorganisms are known to synthesize and accumulate various PHA. Most Pseudomonas strains are able to accumulate PHA. It has been previously demonstrated that from two Iranian isolates of Pseudomonas aeroginosa (PTCC 1310 and PTCC 1740) only the PTCC 1310 contains pha synthase genes. The aim of this study was to clone phaCi and phaC2 genes, which code for PHA synthase enzymes, from P.aeroginosa PTCC1310 into pTZ cloning vector.Materials and methods: Suitable primers were designed according to PHA genomic sequences of Pseudomonas aeroginosa PAO1. The phaCi and phaC2 genes were amplified using PCR technique. The correctness of obtained fragments was confirmed using RsaI and BamHI restriction endonucleases. The PCR products were ligated to pTZ57R cloning vector and used for transformation of competent cells. In order to confirm the correctness of cloning the resulted recombinant plasmids were examined by different restriction endonuclease like Rsai, BamHI. Finally the cloned genes were sequenced and compared with the genes of Pseudomonas aeroginosa PAO 1.Results: Gel electrophoresis of PCR products revealed the expected bands. Restriction digestion of PCR products confirmed the correctness of PCR products. Cultivation of transformed bacteria showed many colonies. Digestion of prepared plasmids with the restriction endonucleases confirmed the correctness of cloning. The comparison of the sequence of the cloned genes with the template genes revealed a high degree of similarity.Discussion: For the first time phaCi and phaC2 genes from an Iranian isolate of Pseudomonas aeroginosa were cloned. The comparison of the cloned genes with the genes of Pseudomonas aeroginosa PA01 revealed a high degree of similarity. Keywords: PHA, phaCi, phaC2, Pseudomonas aeroginosa PTCC1310.
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| Descriptor
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1. Pseudomonas aeruginosa.- Descriptors: Pseudomonas aeruginosa
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Acyltransferases
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Genes
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Polymers
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Enzymes
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Polymerase Chain Reaction
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| Added Entry
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Abedi, Daryoush, Supervisor
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Mir Mohammad Sadeghi, Hamid, Supervisor
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Vallian, Sadeg, Supervisor
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http://elib.mui.ac.ir/site/catalogue/102614
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