Dissertation > Excellent graduate degree dissertation topics show

Cloning Andanalysis of NAC Transcription Factor Gene in Tobacco(Nicotiana Tabacum L.)

Author: WangFeiFei
Tutor: LiuWeiQun
School: Henan Agricultural University
Course: Biochemistry and Molecular Biology
Keywords: Tobacco (Nicotiana tabacum L.) NAC transcription factor Gene clone Expression
CLC: Q943.2
Type: Master's thesis
Year: 2011
Downloads: 14
Quote: 0
Read: Download Dissertation

Abstract


The nicotine is a secondary metabolite synthesized in the root,a tobacco-specific alkaloid. After topping, the content of nicotine accumulated, but the reason why the content increased and the regulation mechanism of nicotine are not clear. Therefore, suppression subtractive hybridization was adapted to construct a cDNA library of root tip before and after topping and screen differentially expressed genes, which has great potential to reveal the molecular basis referred to the impact of topping on the nicotine content.There are a large number of transcription factor genes in plant and they play an important role in regulating plant development and stress response. AP2/EREBP、bZIP、MYB、WRKY和NAC are common transcription factors family. In this study, NAC transcription factor EST sequences was screened out from SSH-cDNA library and it was as a probes to clone the full-length cDNA sequences of NtNAC-R1.It had an important significance on the study of molecular mechanisms in regulation of nicotine biosynthesis. The main results are as follow:(1) Real-time PCR technique was applied to verify expression patterns before and after topping of EST sequences of different gene.It showed that F-box、MYB、NAC、SAHH、ST1 had an increased expression level at 24h after topping,WRKY、AUXIN -induced mRNA had an decreased expression level at 24h after topping.(2) The full-length cDNA sequence of NtNAC-R1 was amplified by in silico cloning and RT-PCR. NtNAC-R1 had an 936bp open reading frame in length, encoding 311 amino acids, with a typical conserved domain of NAC transcription factor family.(3) Tissue-specific analysis showed that the two genes had a high expression level in roots. The expression pattern of NtNAC-R1 in root before and after tobacco topping was analyzed by RT-PCR and Northern blotting. It showed that NtNAC-R1 had a decreased expression level at 2h and 4h after topping, then rise.(4) The recombinant expression vectors of pRSETB-NtNAC-R1 was constructed and transformed into E. coli. The target proteins was successfully expressed in inclusion bodies.(5) The sense and anti-sense expression vectors of NtNAC-R1 gene were constructed and got antisense transgenic plants. The expression patterns of nicotine biosynthesis genes PMT and ODC in antisense transgenic plants were analyzed by RT-PCR. It showed that they had a decreased expression level.

Related Dissertations

  1. Research on Combinatorial Regulation of Multiple Transcription Factors,Q78
  2. The Research of the High-Rise Building’s Form and Expression in the New Century,TU971
  3. Research on Design of Teaching Buildings for Arts Departments of Higher Learning Institutions,TU244.3
  4. Research on Temporal Information Recognition and Normalization,TP391.1
  5. Expression of D-AtCGS in E. Coli and Preparation of Polyclonal Antibody Against D-AtCGS,Q943.2
  6. The Study on Arabidopsis Thaliana Heat Shock Factor HSFA 1d Response to Formaldehyde Stress,Q945.78
  7. Expression of hBMP4 and hBMP7 in Chinese Hamster Ovary Cells,Q78
  8. Cloning and Expression Analysis of GPx, GST and SAHH Genes in Chlamydomonas Sp. ICE-L from Antarctica,Q943.2
  9. cDNA Cloning, Expression of vp5 and vp7 Genes and Subcecullar Localization of VP5 and VP7 Proteins in Grass Carp Reovirus,S941.41
  10. Cloning, Expression of vp6 and ns38 Genes and Immunogenicity of VP6 and NS38 in Grass Carp Reovirus,S941.41
  11. Expression of Prolyl Isomerase Pin1 in Osteosarcoma and the Effect of Regulation on Cell Cycle,R738.1
  12. The Expression Survey of BMP Signalling Pathway in the Human Embryonic Tooth Germ,R78
  13. Preliminary Study of Phosphorylation Effect on UGT1A3 Metabolism Activity Towards Drugs,R346
  14. Cloning and Expression of Growth Hormone Gene in Cobia (Rachycentron Canadus),Q786
  15. Cloning and Characterization of BAFF Gene from Whitespotted Catshark and Crucian Crap,S917.4
  16. Optimization of Fermentation Conditions, Purification, Cloning and Expression of a Cold-active Lipase from Pseudomonas Sp.RT-1,TQ925
  17. Functional Analysis of Proteins Encoded by RNA2 of Wheat Yellow Mosaic Virus,S435.121
  18. Comparative Study on Reproductive Biological Characteristics of Helicoverpa Armigera and Helicoverpa Assulta (Lepidoptera:Nuctuidae),S433
  19. Cloning, Expression and Polymorphism Analysis of BMP4 Gene in Hu Sheep,S826
  20. GIH cDNA Cloning and Mrna Expression in Litopenaeus Vannamei,S917.4
  21. Cloning and Expression Analysis of Scavenger Receptor Class B Type Ⅰ and Antifreeze Proteinstype Ⅱ Genes in Lutjanus Sanguineus,S917.4

CLC: > Biological Sciences > Botany > Plant Cell Genetics > Plant Genetic Engineering
© 2012 www.DissertationTopic.Net  Mobile