Dissertation > Excellent graduate degree dissertation topics show

Study on Constructing Engineered Bacillus Thuringiensis of Efficiently Expressing Tobacco Chitinase Gene TchiB and Insecticidal Activity

Author: LiYuCheng
Tutor: XiaLiQiu
School: Hunan Normal University
Course: Microbiology
Keywords: Bacillus thuringiensis Red/ET homologous recombination site-specific intergration homologous recombination tchiB gene
CLC: S482.3
Type: Master's thesis
Year: 2012
Downloads: 78
Quote: 0
Read: Download Dissertation

Abstract


Bacillus thuringinsis is a kind of environment friendly microbiology agriculture pestcide, which widely used for biological control of farming and forestry pests. The research about Bt engineered strains in domestic and overseas all focused on getting optimized strains which have more better charactetities such as expanding insecticidal spectrum and high toxicity. It was reported that chitinase could increase the insecticidal activity of insecticidal crystal protein markedly by destorying the peritriphic menbrane.We employed homologous recombination' dissertation">Red/ET homologous recombination technology, putting the tobacco Chitinase gene between the cry2Aa promoter gene and cryl Ac terminator gene, to obtain mosaic gene pcry2Aa-tchiB-1Act. Then cloning pcry2Aa-tchiB-1Act gene into shuttle vecter pHT315, constructed recombination plasmid pHTtchiB. Electricing tansform pHTtchiB into Bt subsp.kurstaki HD73strain, got engineered strain HDtchiB. By means of SDS analysis and detection, we found hetero-logous gene tchiB could efficiently expressed under the control of cry2Aa strong promoter, and the protein were further identified by mass spectrometry. The insecticidal activity of the engineered strain shows higher toxicity to Helicoverpa armigera larvae compared with that of original strain HD73.Heterologous gene always is unstabitily or even losed in Bt eng in-eered strains. In order to keep it stability, it is a effective way to integrate heterologous gene into the chrosome of B. thuringinsis. The Chitinase gene located on the chromosome of B. thuringinsis was choosed as the integration site for heterologous gene, then an integrative plasmid pKCHIUN was constructed by cloning the homologous arms into a temperature-sensitive E.coli-Bt shutter vector pKSV7. Employing Red/ET homologous recombination technology constructed a integrative vector pKTCHIUN contained mosaic gene pcry2Aa-tchiB-1Act.Via Red/ET homologous recombination technology constructed the shuttle expression vector pHTtchiB and intergrative vector pKTCHIUN successfully, and then achieved to construct the B.thuringinsis engineered strain HDtchiB that could efficiently express the heterologous gene, which lay an important foundation for studying the exogeneous insecticidal genes that have synergistic effect into B.thuringinsis, and the constructing of engineered strains with superior quality.

Related Dissertations

  1. Insecticidal Activities of Recombinant CryIAc Protein from Bacillus Thuringiensis and Solid Fermentation Products from Aspergillus Terreus PNR2,S476.1
  2. Select of Biofilm Formation Mutants and Cloning and Primary Function Analysis of tatC Gene from Pseudomonas Fluorescens Strain 7-14,S432.4
  3. Preliminary Study on the Homologous Recombination System Mediated by SPP1 Phage Recombinase in Bacillus Subtilis,Q78
  4. Isolation and Molecular Identification of Bacillus Thuringiensis Against Plutella Xylostella,S476.12
  5. Thienamycin biosynthetic gene cluster heterologous expression and transformation,R914
  6. Sequencing of GspF and GspE Gene of Type Ⅱ Serection System in Stenotrophomonas Maltophilia D2 and Constructing of the Recombinant Suicide Plasmid of Smp Gene,Q78
  7. Constrction and Biological Characteristic Identification of Goatpox Virus Gene Deleted Mutant,S852.65
  8. Identifying and Clonging the cry30Ga2 Gene from B. Thuringiensis Isolate S2160-1,Q78
  9. Modification of Phosphoenolpyruvate: Carbohydrate Phosphotransferase System of Escherichia Coli by Genetic Engineering and Characterization of Mutants,Q78
  10. Deregulation of Gene Responsible for Xylose Metabolism in Bacillus Subtilis and Its Phenotypic Effects,Q78
  11. Increase of the Yield of γ-decalactone by Genetically Modified Strain of Yarrowia Lipolytica,TS202.3
  12. The Effect of Astaxanthin Yield by Overexpressing GGPP Synthase in Phaffia Yeast,TQ929
  13. Lidamycin a cloning the bacteria atrA homologous genes and its function,TQ465
  14. Identification of Cry Type Genes from Bacillus Thuringiensis Strains,S476.1
  15. Construction of Mouse HSP10 SiRNA and Overexpression Adenovirus Vector and Investigation of Its Role on the Apoptosis of Mouse Ovarian Granulosa Cells,R363
  16. Study on ALD4 Gene Disruption and GPD1 Silencing in Saccharomyces Cerevisiae Industrial Strain Y01,Q78
  17. Construction of Recombinant Adenovirus Vector Co-expressing P12A and 3C Genes of FMDV O Virus,S852.65
  18. Breeding and Fermentation Conditions Optimization of Associated Bacteria in 2-keto-L-Gulonic Acid Fermentation,TQ921
  19. Screening and identification of one pair the tobacco A high virulence of Bacillus thuringiensis,S476.1
  20. Cloning, Expression and Insecticidal Activity of Novel cry9 Genes from Bacillus Thuringiensis,S476.1
  21. Regulation and Expression of gab Gene Cluster in Bacillus Thuringiensis,Q78

CLC: > Agricultural Sciences > Plant Protection > Pesticide ( chemical control ) > Various pesticides > Pesticides
© 2012 www.DissertationTopic.Net  Mobile