Dissertation > Excellent graduate degree dissertation topics show

Localization of the Protein Encoded by a Gene that is Closely Related to the Synthesis of Magnetosome in Magnetospirillum Gryphiswaldense MSR-1

Author: ChenQi
Tutor: LiFeng
School: Huaibei Normal
Course: Botany
Keywords: magnetotactic bacteria Magnetospirillum gryphiswaldense MSR-1 magnetosome localization
CLC: Q936
Type: Master's thesis
Year: 2011
Downloads: 48
Quote: 0
Read: Download Dissertation

Abstract


Magnetospirillum gryphiswaldense MSR-1, the modal strain of genus Magnetospirillum, is capable of synthesizing specific intracellular structures, magnet- osomes, which are chain-linked and membrane-enclosed crystals of magnetic iron oxide. At the beginning of our experiments, a magnetosome-deleted mutant NM21 of Magnetospirillum gryphiswaldense MSR-1 was generated by mini-Tn5 lacZ2 transposon mutagenesis, and a 3073-bp fragment flanking mini-Tn5 lacZ2 in NM21 was cloned by Anchored PCR. Sequencing analysis showed that this fragment contained three putative ORFs. Analysis of the protein encoded by ORF1 suggested that it probably located on the membrane of magnetosomes, take part in the biosyn- thesis of magnetosomes as Fe2+ transporter. In confirmation of the hypothesis, the subcellular localization of a protein encoded by ORF1 from 3073-bp fragment in Magnetospirillum gryphiswaldense MSR-1 was studied.Analysis of some bio-software predicted that ORF1 encoded a putative protein of 320 amino acids. There were no signal peptide in protein ORF1 which contained 5 hydrophobic regions, 4 transmembrane domains, 18 protein phosphorylation sites, 8 -helixes and 2β-sheets, most probably located on plasma membrane or extracellular. These informations will help us to study the ORF1 protein.The ORF1 was PCR amplified with genomic DNA of MSR-1 as a template, the EGFP was amplified from plasmid pEGFPN-1 respectively; The ORF1‘was PCR am- plified with genomic DNA of MSR-1 as a template, the EGFP‘was amplified from plasmid pEGFPN-1 respectively. Both of them were subcloned into the broad- host-range vector pBBR1MCS-2, then transformed into E. coli S17-1 to generate fusion expression vector pBBR1MCS-ORF1-EGFP and pBBR1MCS- EGFP‘-ORF1‘. They were transferred into MSR-1 by conjugation from E. coli S17-1. Localization of ORF1 protein was examined by fluorescence microscopy after the positive clone were induced for protein expression with a result of on the membrane of magnetosomes.The ORF1‘was PCR amplified with genomic DNA of M. gryphiswaldense as a template then cloned into the pET-29a vector. The recombinant plasmid pET-29a- ORF1‘was transformed into E. coli BL 21 and induced for expression. SDS-PAGE profile showed there was no clear increased protein band.Research preliminarily demonstreted that the protein encoded by ORF1 precisely located on the membrane of magnetosomes. The result laid a foundation for the research of its function, as well as the further research of the role of the 3073-bp and its flanking fragment in the biosynthesis of magnetosome in MSR-1.

Related Dissertations

  1. Technology for Localization Attack Detection in Wireless Sensor Networks,TP212.9
  2. cDNA Cloning, Expression of vp5 and vp7 Genes and Subcecullar Localization of VP5 and VP7 Proteins in Grass Carp Reovirus,S941.41
  3. Large the Hongshan iron ore mine personnel tracking positioning system optimization study,TN929.5
  4. Interpretation of Chinese TV Marriage Program under the Influence of Consumer Culture,G222.3
  5. A Study on the Development of the Resources Localilzation of the Middle School Art Teaching in Heyuan,G633.955
  6. Genetic Model Analysis and QTL Mapping of Agronomic and Quality Traits in Soybean,S565.1
  7. Functional Analysis of Proteins Encoded by RNA2 of Wheat Yellow Mosaic Virus,S435.121
  8. Indirect Imaging of Singlet Oxygen Generation from a Single Cell,Q2-3
  9. Expressive Characterization of Transcription Factors in Cotton Fibre Initiation and Cloning and Characterization of Three Transcription Factor Genes in Gossypium Hirsutum L.,Q943
  10. Cloning and Characterization of Photoperiod Sensitive Gene ZmELF4 in Maize,S513
  11. Establishment of ELISA Based on Recombinant Protein NS1 of Swine Influenza Virus and Nuclear Localization of SIV NS and NP Proteins,S858.28
  12. Research and Design on Indoor Localization System Based on ZigBee,TN929.5
  13. The Research on Localization Algorithm of Wireless Sensor Networks,TN929.5
  14. Expression Analysis and Localization of OsDMI3 Gene Induced by Abscisic Acid,S511
  15. Cloning and Expressing Profiles of Eimeria Tenella Rhoptry Neck Protein 2 EtRON2 Gene during the Different Development Stages,S852.7
  16. Silkworm circadian clock gene cloning and functional characterization Cry,S881.2
  17. Folk Art and Modern Mix of Advertising Research,J524.3
  18. Project planning and site selection and engineers from the perspective of values ??in our localization Engineering Ethics,B82-057
  19. The Research on the Lucky Pattern of Folk Residences and Furniture of Western Hunan,J525
  20. A Comparative Research on the Development of Modern Sino-US Public Administration,D035
  21. On the Localization of Logo Design in the Modern Role,J524

CLC: > Biological Sciences > Microbiology > Microbial biochemistry
© 2012 www.DissertationTopic.Net  Mobile