DOCSLIB.ORG

(S)-Stylopine Synthase and Two FAD Oxidases from a Cdna Library from Argemone Mexicana

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
(S)-Stylopine Synthase and Two FAD Oxidases from a Cdna Library from Argemone Mexicana Isolation and characterization of (S)-chelianthifoline synthase, (S)-stylopine synthase and two FAD oxidases from a cDNA library from Argemone mexicana Dissertation zur Erlangung des akademischen Grades doctor rerum naturalium (Dr. rer. nat.) vorgelegt an der Naturwissenschaftlichen Fakultät I-Biowissenschaften der Martin-Luther-Universität Halle-Wittenberg Fachbereich Biochemie / Biotechnologie von Frau María Luisa Díaz Chávez geboren am 25.03.1974 in Mexiko Gutachter: 1. Prof. Dr. Toni M. Kutchan 2. Prof. Dr. Jörg Degenhardt 3. Prof. Dr. Ute Wittstock Halle, den 22. Januar 2009 urn:nbn:de:gbv:3-000015158 [http://nbn-resolving.de/urn/resolver.pl?urn=nbn%3Ade%3Agbv%3A3-000015158] This work was supported by DAAD/CONACyT and PROMEP/UAEM Content Content CONTENT……………………………………………………………………………………. I ABBREVIATIONS ………………………………………......………...………………….. IV 1. INTRODUCTION ..................................................................................................1 1.1. Alkaloids.................................................................................................................... 1 1.1.1. Benzylisoquinoline alkaloids ................................................................................. 1 1.1.2. Biosynthesis of berberine and sanguinarine........................................................... 3 1.2. Cytochrome P450..................................................................................................... 4 1.2.1. Plant cytochrome P450........................................................................................... 5 1.3. Berberine bridge enzyme (BBE) and BBE-like proteins ...................................... 7 1.4. (S)-tetrahydroprotoberberine oxidase (STOX) ..................................................... 9 1.5. Argemone mexicana L. ........................................................................................... 10 2. MATERIAL.........................................................................................................12 2.1. Enzymes................................................................................................................... 12 2.2. Proteins.................................................................................................................... 12 2.3. Nucleotides .............................................................................................................. 12 2.4. DNA fragments....................................................................................................... 12 2.5. Cloning vectors....................................................................................................... 12 2.6. Synthetic Oligonucleotides..................................................................................... 12 2.7. Organisms ............................................................................................................... 13 2.7.1. Plants .................................................................................................................... 13 2.7.2. Bacteria................................................................................................................. 13 2.7.3. Insect cells work................................................................................................... 13 2.8. Antibiotics ............................................................................................................... 13 2.9. Internet searches and alignments ......................................................................... 13 2.10. Chemicals ................................................................................................................ 14 2.11. Kits........................................................................................................................... 15 2.12. Consumables ........................................................................................................... 15 2.13. Instruments ............................................................................................................. 15 3. METHODS..........................................................................................................16 3.1. Alkaloids.................................................................................................................. 16 3.1.1. Alkaloid extraction from plant tissues.................................................................. 16 3.1.2. Analyses by High Performance Liquid Chromatography (HPLC) ...................... 16 3.1.3. Analyses by Liquid Chromatography- Mass Spectrometry (LC-MS, TOF)........ 16 3.2. Isolation of RNA..................................................................................................... 17 3.2.1. RNA Isolation....................................................................................................... 17 3.2.2. Poly-(A)+ RNA isolation ...................................................................................... 18 3.3. Isolation of DNA..................................................................................................... 18 3.3.1. Plasmid DNA purification.................................................................................... 18 3.3.2. Purification of DNA fragments from agarose gel ................................................ 18 3.3.3. Baculovirus DNA from infected Sf9 cells............................................................ 18 3.4. Insect cell culture.................................................................................................... 19 3.4.1. Maintenance ......................................................................................................... 19 3.5. Electrophoresis ....................................................................................................... 19 3.5.1. Protein polyacrylamide gel electrophoresis (PAGE) ........................................... 19 3.5.2. RNA agarose gel .................................................................................................. 20 3.5.3. DNA agarose gel .................................................................................................. 21 I Content 3.6. cDNA library........................................................................................................... 21 3.6.1. λ-cDNA library construction................................................................................ 21 3.7. Preparation of plating cells for library amplification......................................... 22 3.8. Plating bacteriophage ......................................................................................... 22 3.9. Picking bacteriophage plaques........................................................................... 22 3.10. Library screening................................................................................................... 22 3.11. Northern Blot.......................................................................................................... 23 3.11.1. Blotting............................................................................................................. 23 3.11.2. Prehybridization ............................................................................................... 24 3.11.3. Random primer labelling of DNA.................................................................... 24 3.11.4. Hybridization.................................................................................................... 25 3.12. First-strand cDNA synthesis.................................................................................. 25 3.13. Rapid amplification of 5’-cDNA ends (5’-RACE)............................................... 26 3.14. Polymerase chain reaction (PCR)......................................................................... 26 3.14.1. Standard PCR reaction ..................................................................................... 26 3.14.2. Screening bacterial colonies by PCR ............................................................... 27 3.14.3. Sequencing of DNA ......................................................................................... 27 3.15. DNA modifications................................................................................................. 28 3.15.1. Addition of a 3’-A Overhang ........................................................................... 28 3.15.2. Dephosphorylation of DNA fragments ............................................................ 28 3.15.3. Restriction enzyme digestion ........................................................................... 29 3.16. DNA cloning............................................................................................................ 29 3.16.1. TA cloning........................................................................................................ 29 3.16.2. Subcloning........................................................................................................ 29 3.17. Transformation of competent cells ....................................................................... 30 3.18. Protein Expression................................................................................................. 30 3.18.1. BaculoGold Expression Vector System ........................................................... 30 3.18.2. Co-transfection using BD Baculogold ............................................................. 31 3.18.3. BAC-to-BAC expression system.....................................................................
Load more

Recommended publications
  • 1. Introduction
    Introduction 1. Introduction 1.1. Alkaloids The term alkaloid is derived from Arabic word al-qali, the plant from which “soda” was first obtained (Kutchan, 1995). Alkaloids are a group of naturally occurring low-molecular weight nitrogenous compounds found in about 20% of plant species. The majority of alkaloids in plants are derived from the amino acids tyrosine, tryptophan and phenylalanine. They are often basic and contain nitrogen in a heterocyclic ring. The classification of alkaloids is based on their carbon-nitrogen skeletons; common alkaloid ring structures include the pyridines, pyrroles, indoles, pyrrolidines, isoquinolines and piperidines (Petterson et al., 1991; Bennett et al., 1994). In nature, plant alkaloids are mainly involved in plant defense against herbivores and pathogens. Many of these compounds have biological activity which makes them suitable for use as stimulants (nicotine, caffeine), pharmaceuticals (vinblastine), narcotics (cocaine, morphine) and poisons (tubocurarine). The discovery of morphine by the German pharmacist Friedrich W. Sertürner in 1806 began the field of plant alkaloid biochemistry. However, the structure of morphine was not determined until 1952 due to its stereochemical complexity. Major technical advances occurred in this field allowing for the elucidation of selected alkaloid biosynthetic pathways. Among these were the introduction of radiolabeled precursors in the 1950s and the establishment in the 1970s of plant cell suspension cultures as an abundant source of enzymes that could be isolated, purified and characterized. Finally, the introduction of molecular techniques has made possible the isolation of genes involved in alkaloid secondary pathways (Croteau et al., 2000; Facchini, 2001). 1.1.1. Benzylisoquinoline alkaloids Isoquinoline alkaloids represent a large and varied group of physiologically active natural products.
    [Show full text]
  • Influence of Different Elicitors on BIA Production in Macleaya Cordata
    www.nature.com/scientificreports OPEN Infuence of diferent elicitors on BIA production in Macleaya cordata Peng Huang1,2,7, Liqiong Xia3,7, Li Zhou1,7, Wei Liu1,4, Peng Wang1, Zhixing Qing5* & Jianguo Zeng1,6* Sanguinarine (SAN) and chelerythrine (CHE) have been widely used as substitutes for antibiotics for decades. For a long time, SAN and CHE have been extracted from mainly Macleaya cordata, a plant species that is a traditional herb in China and belongs to the Papaveraceae family. However, with the sharp increase in demand for SAN and CHE, it is necessary to develop a new method to enhance the supply of raw materials. Here, we used methyl jasmonate (MJ), salicylic acid (SA) and wounding alone and in combination to stimulate aseptic seedlings of M. cordata at 0 h, 24 h, 72 h and 120 h and then compared the diferences in metabolic profles and gene expression. Ultimately, we found that the efect of using MJ alone was the best treatment, with the contents of SAN and CHE increasing by 10- and 14-fold, respectively. However, the increased SAN and CHE contents in response to combined wounding and MJ were less than those for induced by the treatment with MJ alone. Additionally, after MJ treatment, SAN and CHE biosynthetic pathway genes, such as those encoding the protopine 6-hydroxylase and dihydrobenzophenanthridine oxidase enzymes, were highly expressed, which is consistent with the accumulation of SAN and CHE. At the same time, we have also studied the changes in the content of synthetic intermediates of SAN and CHE after elicitor induction.
    [Show full text]
  • Argemone Ochroleuca: (PAPAVERACEAE), ALKALOID POTENTIAL SOURCE for AGRICULTURAL and MEDICINAL USES †
    Tropical and Subtropical Agroecosystems 23 (2020): #31 Hernández-Ruiz et al., 2020 Review [Revisión] Argemone ochroleuca: (PAPAVERACEAE), ALKALOID POTENTIAL SOURCE FOR AGRICULTURAL AND MEDICINAL USES † [Argemone ochroleuca: (PAPAVERACEAE), FUENTE POTENCIAL DE ALCALOIDES PARA LA AGRICULTURA, Y USO MEDICINAL] J. Hernández-Ruiz1, J. Bernal2, J. Gonzales-Castañeda1, J. E. Ruiz-Nieto1 and A. I. Mireles-Arriaga1* 1División de Ciencias de la Vida, Universidad de Guanajuato. Km 9 carretera Irapuato-Silao, Ex Hacienda. El Copal, Irapuato, Guanajuato. 36500 México. Email: [email protected] 2Department of Entomology, Texas A&M University, College Station, TX 77843-247, USA *Corresponding author SUMMARY Background. The genus Argemone contains 24 species, A. ochorleuca is present in national territory and is used in agriculture and traditional medical treatments for various conditions. Results. A. ochorleuca is an herbaceous and/or perennial plant that blooms all year. This plant had the potential as a source of benzyl isoquinoline alkaloids, which are the main bioactive compounds responsible for antibacterial, antifungal properties. However, some of these compounds are associated with toxic effects too. Information about concentrations and parts of the plant it is important for all uses and applications. Implications. The present work summarizes available information on phytochemical and medicinal properties. Conclusion. In A. ochrolecuca, six of the 45 alkaloids reported for the genus Argemone have been studied, dihydro-keleritrin and dihydro-sanguiranine are the most abundant in the seeds and vegetative tissue of the species. The updated information should be useful to guide future research on this plant. Keywords: Alkaloids; papaveraceae; berberine; sanguinarine. RESUMEN Antecedentes. El género Argemone contiene 24 especies, A.
    [Show full text]
  • S41438-020-00450-6.Pdf
    Xu et al. Horticulture Research (2021) 8:16 Horticulture Research https://doi.org/10.1038/s41438-020-00450-6 www.nature.com/hortres ARTICLE Open Access Integration of full-length transcriptomics and targeted metabolomics to identify benzylisoquinoline alkaloid biosynthetic genes in Corydalis yanhusuo Dingqiao Xu1,HanfengLin 2,YupingTang1,LuHuang1,JianXu2,SihuiNian3 and Yucheng Zhao 2 Abstract Corydalis yanhusuo W.T. Wang is a classic herb that is frequently used in traditional Chinese medicine and is efficacious in promoting blood circulation, enhancing energy, and relieving pain. Benzylisoquinoline alkaloids (BIAs) are the main bioactive ingredients in Corydalis yanhusuo. However, few studies have investigated the BIA biosynthetic pathway in C. yanhusuo, and the biosynthetic pathway of species-specific chemicals such as tetrahydropalmatine remains unclear. We performed full-length transcriptomic and metabolomic analyses to identify candidate genes that might be involved in BIA biosynthesis and identified a total of 101 full-length transcripts and 19 metabolites involved in the BIA biosynthetic pathway. Moreover, the contents of 19 representative BIAs in C. yanhusuo were quantified by classical targeted metabolomic approaches. Their accumulation in the tuber was consistent with the expression patterns of identified BIA biosynthetic genes in tubers and leaves, which reinforces the validity and reliability of the analyses. Full- length genes with similar expression or enrichment patterns were identified, and a complete BIA biosynthesis pathway fi 1234567890():,; 1234567890():,; 1234567890():,; 1234567890():,; in C. yanhusuo was constructed according to these ndings. Phylogenetic analysis revealed a total of ten enzymes that may possess columbamine-O-methyltransferase activity, which is the final step for tetrahydropalmatine synthesis. Our results span the whole BIA biosynthetic pathway in C.
    [Show full text]
  • Plant Profile, Phytochemistry and Pharmacology of Argemone Mexicana Linn
    Innovare International Journal of Pharmacy and Pharmaceutical Sciences Academic Sciences ISSN- 0975-1491 Vol 6, Issue 7, 2014 Review Article PLANT PROFILE, PHYTOCHEMISTRY AND PHARMACOLOGY OF ARGEMONE MEXICANA LINN. A REVIEW REKHA SHARANAPPA, VIDYASAGAR G. M.* Medicinal Plants and Microbiology Research Laboratory, Department of Post-Graduate Studiesand Research in Botany, Gulbarga University, Gulbarga-585106, Karnataka, India. Email: [email protected] Received: 02 June 2014 Revised and Accepted: 13 Jul 2014 ABSTRACT Argemone Mexicana is extensively used as traditional medicine for the treatment of numerous diseases. Various parts of the plant were extensively used in Ayurveda, Siddha, Unani and Homeopathic medicines. It is reported to have antimicrobial activity, wound healing property, larvicidal and chemosterilant activity, nematicidal and allelopathic potential, antimalarial, antibacterial and antifungal, molluscicidal, anticancer, hepatoprotective, anti-HIV and neuropharmacological activity. Chemical investigations of this plant have revealed the presence of alkaloids, amino acids, phenolics and fatty acids. A. mexicana has shown promise as an effective bio-control agent. The present review includes the detailed exploration of traditional uses, phytochemical and pharmacological properties and actions of whole plant extract reported so far. Keywords: Plant Profile, Phytochemistry, Pharmacology, Biological activity, Argemone mexicana, Toxicity, Safety Evaluation. INTRODUCTION review and compile updated information on various aspects of A. mexicana, a plant used all over the world. Plants have been used in medicines since time immemorial. India has a rich heritage of using medicinal plants in traditional medicines, Habitat as in the Ayurveda, Siddha and Unani systems besides folklore practices. The earliest inscription of the medicinal uses of plants is A. mexicana is native of tropical America which has distributed in found in the “Rigveda”, which is one of the oldest repositories of tropical and subtropical regions of the World.
    [Show full text]
  • Considérations Sur L'histoire Naturelle Des Ranunculales
    Considérations sur l’histoire naturelle des Ranunculales Laetitia Carrive To cite this version: Laetitia Carrive. Considérations sur l’histoire naturelle des Ranunculales. Botanique. Université Paris-Saclay, 2019. Français. NNT : 2019SACLS177. tel-02276988 HAL Id: tel-02276988 https://tel.archives-ouvertes.fr/tel-02276988 Submitted on 3 Sep 2019 HAL is a multi-disciplinary open access L’archive ouverte pluridisciplinaire HAL, est archive for the deposit and dissemination of sci- destinée au dépôt et à la diffusion de documents entific research documents, whether they are pub- scientifiques de niveau recherche, publiés ou non, lished or not. The documents may come from émanant des établissements d’enseignement et de teaching and research institutions in France or recherche français ou étrangers, des laboratoires abroad, or from public or private research centers. publics ou privés. Considérations sur l’histoire naturelle des Ranunculales 2019SACLS177 Thèse de doctorat de l'Université Paris-Saclay : préparée à l’Université Paris-Sud NNT École doctorale n°567 : Sciences du végétal, du gène à l'écosystème (SDV) Spécialité de doctorat : Biologie Thèse présentée et soutenue à Orsay, le 05 juillet 2019, par Laetitia Carrive Composition du Jury : Catherine Damerval Directrice de recherche, CNRS (– UMR 320 GQE) Présidente du jury Julien Bachelier Professeur, Freie Universität Berlin (– Institute of Biology) Rapporteur Thomas Haevermans Maître de conférences, MNHN (– UMR 7205 ISYEB) Rapporteur Jean-Yves Dubuisson Professeur, SU (–UMR 7205 ISYEB) Examinateur Sophie Nadot Professeure, U-PSud (– UMR 8079 ESE) Directrice de thèse « Le commencement sera d’admirer tout, même les choses les plus communes. Le milieu, d’écrire ce que l’on a bien vu et ce qui est d’utilité.
    [Show full text]
  • Modulation of Benzylisoquinoline Alkaloid Biosynthesis by Heterologous Expression of Cjwrky1 in Eschscholzia Californica Cells
    RESEARCH ARTICLE Modulation of benzylisoquinoline alkaloid biosynthesis by heterologous expression of CjWRKY1 in Eschscholzia californica cells Yasuyuki Yamada¤*, Tomoe Shimada, Yukiya Motomura, Fumihiko Sato* Department of Plant Gene and Totipotency, Division of Integrated Life Science, Graduate School of Biostudies, Kyoto University, Kyoto, Japan ¤ Current address: Laboratory of Medicinal Cell Biology, Kobe Pharmaceutical University, Kobe, Japan * [email protected] (YY); [email protected] (FS) a1111111111 a1111111111 a1111111111 a1111111111 Abstract a1111111111 Transcription factors control many processes in plants and have high potentials to manipu- late specialized metabolic pathways. Transcriptional regulation of the biosynthesis of mono- terpenoid indole alkaloids (MIAs), nicotine alkaloids, and benzylisoquinoline alkaloids (BIAs) has been characterized using Catharanthus roseus, Nicotiana and Coptis plants. However, OPEN ACCESS metabolic engineering in which specific transcription factors are used in alkaloid biosynthe- Citation: Yamada Y, Shimada T, Motomura Y, Sato sis is limited. In this study, we characterized the effects of ectopic expression of CjWRKY1, F (2017) Modulation of benzylisoquinoline alkaloid which is a transcriptional activator with many targets in BIA biosynthesis in Coptis japonica biosynthesis by heterologous expression of CjWRKY1 in Eschscholzia californica cells. PLoS (Ranunculaceae) and Eschscholzia californica (California poppy, Papaveraceae). Heterolo- ONE 12(10): e0186953. https://doi.org/10.1371/ gous expression of CjWRKY1 in cultured California poppy cells induced increases in tran- journal.pone.0186953 scripts of several genes encoding BIA biosynthetic enzymes. Metabolite analyses indicated Editor: Turgay Unver, Dokuz Eylul Universitesi, that the overexpression of the CjWRKY1 gene also induced increases in the accumulation TURKEY of BIAs such as sanguinarine, chelerythrine, chelirubine, protopine, allocryptopine, and 10- Received: June 30, 2017 hydroxychelerythrine in the culture medium.
    [Show full text]
  • Characterization of Sanguinarine Reductases from Papaver Somniferum (Unpublished Master's Thesis)
    University of Calgary PRISM: University of Calgary's Digital Repository Graduate Studies The Vault: Electronic Theses and Dissertations 2015-05-20 Characterization of sanguinarine reductases from Papaver somniferum Bross, Crystal Bross, C. (2015). Characterization of sanguinarine reductases from Papaver somniferum (Unpublished master's thesis). University of Calgary, Calgary, AB. doi:10.11575/PRISM/25290 http://hdl.handle.net/11023/2260 master thesis University of Calgary graduate students retain copyright ownership and moral rights for their thesis. You may use this material in any way that is permitted by the Copyright Act or through licensing that has been assigned to the document. For uses that are not allowable under copyright legislation or licensing, you are required to seek permission. Downloaded from PRISM: https://prism.ucalgary.ca UNIVERSITY OF CALGARY Characterization of sanguinarine reductases from Papaver somniferum by Crystal Dawn Bross A THESIS SUBMITTED TO THE FACULTY OF GRADUATE STUDIES IN PARTIAL FULFILMENT OF THE REQUIREMENTS FOR THE DEGREE OF MASTER OF SCIENCE DEPARTMENT OF BIOLOGICAL SCIENCES CALGARY, ALBERTA MAY, 2015 © Crystal Dawn Bross 2015 Abstract Papaver somniferum (opium poppy) produces several pharmacologically relevant benzylisoquinoline alkaloids, such as the analgesics codeine and morphine, the muscle relaxant papaverine, the potential anti-cancer drug noscapine, and the antimicrobial agent sanguinarine. Sanguinarine is a highly cytotoxic benzophenanthridine alkaloid synthesized by the plant to defend against herbivory and pathogens. However, sanguinarine can bind DNA, induce apoptosis, and will inhibit the growth of plant cell cultures that do not synthesize benzophenanthridine alkaloids. Therefore, it is proposed that sanguinarine reductase (SanR) exists in plants that synthesize benzophenanthridine alkaloids to facilitate the detoxification of sanguinarine through its reduction to dihydrosanguinarine.
    [Show full text]
  • Modulation of Benzylisoquinoline Alkaloid Biosynthesis By
    www.nature.com/scientificreports OPEN Modulation of benzylisoquinoline alkaloid biosynthesis by overexpression berberine bridge Received: 20 July 2018 Accepted: 11 November 2018 enzyme in Macleaya cordata Published: xx xx xxxx Peng Huang1,2, Wei Liu1,3, Min Xu1,2, Ruolan Jiang1,2, Liqiong Xia4, Peng Wang1, Han Li5, Zhaoshan Tang6, Qiyin Zheng4 & Jianguo Zeng1,7 Macleaya cordata produces a variety of benzylisoquinoline alkaloids (BIAs), such as sanguinarine, protopine, and berberine, which are potential anticancer drugs and natural growth promoters. The genes encoding the berberine bridge enzyme (BBE) were isolated from M. cordata and Papaver somniferum, and then the two genes were overexpressed in M. cordata. Through liquid chromatography with triple-quadrupole mass spectrometry analysis, it was determined that McBBE-OX caused higher levels of (S)-norcoclaurine, (S)-coclaurine, (S)-N-cis-methylcoclaurine, (S)-reticuline, (S)- tetrahydrocolumbamine, (S)-tetrahydroberberine, (S)-cheilanthifoline, and (S)-scoulerine than PsBBE-OX, empty vector or control treatments. qRT-PCR analysis demonstrated that the introduced genes in the transgenic lines were all highly expressed. However, the levels of sanguinarine (SAN) and chelerythrine (CHE) in all the transgenic lines were slightly lower than those in the wild-type lines, possibly because the overexpression of McBBE causes feedback-inhibition. This is the frst report on the overexpression of potential key genes in M. cordata, and the fndings are important for the design of metabolic engineering
    [Show full text]
  • 5. Discussion
    Discussion 5. Discussion 5.1. Argemone mexicana Previous studies on Argemone mexicana, a medicinal plant used for the treatment of pain, diarrhea and cancer, have shown the presence of alkaloids belonging to various classes such as protopines, protoberberines and benzophenanthridine type alkaloids; some of them with antimicrobial and antiallergenic activity. In this work, alkaloid analysis in A. mexicana quantified protopine and allocryptopine as the main compounds in the plant being protopine accumulation three times higher in roots than in leaves. Berberine, reticuline, cryptopine, dihydrosanguinarine and chelerythrine were also found in roots, leaves and stem and sanguinarine was mainly found in roots otherwise only traces were found. The construction of a cDNA library from Argemone mexicana roots generated approximately 160,000 cDNAs. About 1600 randomly selected cDNAs were partially sequenced from the 5- end and the thus generated expressed sequence tags (5-ESTs) were clustered into 1255 unique sequences. Similarity search of the non-redundant ESTs against the public non- redundant database at the NCBI indicated that 43% of these sequences displayed similarity to genes of known function, 36% to hypothetical genes, and the remaining 21% were novel sequences. ESTs were classified into 7 different categories according to putative protein function. To our knowledge, this is the first report of a cDNA library from Argemone mexicana and therefore it may represent a valuable source for gene discovery. In addition, benzylisoquinoline alkaloid biosynthesis in this plant can now potentially be modified, since a transformation protocol using Agrobacterium tumefaciens is established (Godoy-Hernandez et al., 2008). Analysis of the ESTs lead to the identification of several clones homologous to methyltransferases, P450 monooxygenases and FAD dependent oxidoreductases from the biosynthesis of benzylisoquinoline alkaloids.
    [Show full text]
  • FROM Aristolochia Fimbriata, a BASAL ANGIOSPERM
    SEQUENCING AND ANALYSIS OF A cDNA ENCODING A PUTATIVE COCLAURINE N-METHYLTRANSFERASE (CNMT) FROM Aristolochia fimbriata, A BASAL ANGIOSPERM DISSERTATION Submitted To THE FACULTY OF LIFE & ENVIRONMENTAL SCIENCES UNIVERSITY OF PESHAWAR In fulfillment for the degree Of DOCTOR OF PHILOSOPHY by ROSHAN ALI THE CENTER OF BIOTECHNOLOGY AND MICROBIOLOGY UNIVERSITY OF PESHAWAR 2010 1 CENTER OF BIOTECHNOLOGY AND MICROBIOLOGY UNIVERSITY OF PESHAWAR SEMESTER 2010 This Dissertation is Submitted By ROSHAN ALI Titled SEQUENCING AND ANALYSIS OF A cDNA ENCODING A PUTATIVE COCLAURINE N-METHYLTRANSFERASE (CNMT) FROM Aristolochia fimbriata, A BASAL ANGIOSPERM In fulfillment of the requirements for the degree Of DOCTOR OF PHILOSOPHY In BIOTECHNOLOGY 2 APPROVAL The dissertation of Mr. Roshan Ali is approved SUPERVISOR: Dr. Ghosia Lutfullah Signature:______________ Professor, Date:__________________ Center of Biotechnology & Microbiology, University of Peshawar. EXAMINER: Signature:_______________ Date:___________________ 3 4 In the name of Allah, the Beneficent, the Merciful 5 Graphical Abstract of the dissertation: Image sources are: (http://1.bp.blogspot.com/_vKxDQBwCntA/S-DMy0e3g VI/AAAAAAAAAJo/e2ickYGXEas/s1600/2516243964_5e41ba8d14.jpg), (http://images19.fotki.com/v27/photos/6/642761/4094024/Aristolochiafimbriat a37-vi.jpg?1182176542), (http://www.scripps.edu/chem/wong/PIX/riboso me.jpg) and this dissertation. 6 TABLE OF CONTENTS Title Page ACKNOWLEDGEMENTS I ABSTRACT V LIST OF ABBREVIATIONS VII LIST OF FIGURES IX LIST OF TABLES XII LIST OF APPENDICES XIII 1. INTRODUCTION 1 1.1. S-adenosyl-L-Methionine-Dependent Methyltransferases 3 1.2. S-Adenosyl-L-Methionine (SAM or AdoMet) 4 1.3. Genes of Bezylisoquinoline Alkaloid Pathway 4 1.4. Alkaloids 6 1.4.1.
    [Show full text]
  • Bioactive Alkaloids from Medicinal Plants of Bhutan
    University of Wollongong Research Online University of Wollongong Thesis Collection 1954-2016 University of Wollongong Thesis Collections 2004 Bioactive alkaloids from medicinal plants of Bhutan Phurpa Wangchuk University of Wollongong Follow this and additional works at: https://ro.uow.edu.au/theses University of Wollongong Copyright Warning You may print or download ONE copy of this document for the purpose of your own research or study. The University does not authorise you to copy, communicate or otherwise make available electronically to any other person any copyright material contained on this site. You are reminded of the following: This work is copyright. Apart from any use permitted under the Copyright Act 1968, no part of this work may be reproduced by any process, nor may any other exclusive right be exercised, without the permission of the author. Copyright owners are entitled to take legal action against persons who infringe their copyright. A reproduction of material that is protected by copyright may be a copyright infringement. A court may impose penalties and award damages in relation to offences and infringements relating to copyright material. Higher penalties may apply, and higher damages may be awarded, for offences and infringements involving the conversion of material into digital or electronic form. Unless otherwise indicated, the views expressed in this thesis are those of the author and do not necessarily represent the views of the University of Wollongong. Recommended Citation Wangchuk, Phurpa, Bioactive alkaloids from medicinal plants of Bhutan, M.Sc. thesis, Department of Chemistry, University of Wollongong, 2004. http://ro.uow.edu.au/theses/315 Research Online is the open access institutional repository for the University of Wollongong.
    [Show full text]