Dissertation > Excellent graduate degree dissertation topics show

Cloning and Characterization of BAFF Gene from Whitespotted Catshark and Crucian Crap

Author: PangShuYing
Tutor: RenWenHua
School: Nanjing Normal University
Course: Biochemistry and Molecular Biology
Keywords: Chilosoyllium plagiosum Carassius auratus BAFF clone invitro expression biological information
CLC: S917.4
Type: Master's thesis
Year: 2011
Downloads: 5
Quote: 0
Read: Download Dissertation


This thesis contains four chapters:1. Some advances in the studies of BAFF (B cell activating factor of the TNF family) were reviewed.2. Discussion about the meaning of BAFF gene researches in fishes.3. In this study, a CpBAFF was amplified from Chiloscyllium plagiosum by RT-PCR and RACE strategies. The open reading frame of CpBAFF cDNA consists of 819 bases encoding a protein of 272 amino acids. Like other reported BAFF molecues, CpBAFF includes a typically transmembrane domain and a furin protease cleavage site. Sequence alignment shows that CpBAFF shares 36%-57% identity with BAFF sequences reported in other vertebrate animals, and it shares identical structural features. Real-time quantitative RT-PCR demonstrated high expression of CpBAFF in the main immune organ spleen, moderate expression in kidney, brankia and liver, and low expression in intestine, heart and pancreas. The predicted 3D structural analysis of the soluble mature part of CpBAFF (CpsBAFF) analyzed by comparative protein modeling revealed that it is very similar to the 3D structure of human BAFF. Recombinant CpsBAFF fused with NusA-His6 tag was efficiently expressed in Escherichia coli BL21 (DE3) and its molecular weight of-83 KDa was identified by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and Western blotting. In vitro MTT assay indicated that the purified pET43.1a (+)-CpsBAFF protein was able to promote splenic B cells survival in a dose-dependent manner. The present findings will provide valuable information for the shark immunity and the evolution of vertebrate adaptive immunity.4. In the present study, a CaBAFF gene was amplified from Carassius auratus by RT-PCR and RACE strategies. The full-length cDNA of CaBAFF was 1639 bp, consisiting of a 407 bp 5’-terminal untanslated region (UTR), and a 795 bp open reading frame (ORF) which encods a protein containing 264-amino acids bearing characteristics of the TNF family. Like other reported BAFF molecules, CaBAFF contains a typically transmembrane domain and a TNF homology domain. Sequence alignment shows that CaBAFF shares 47%-77% identity at the amino acid level with BAFF sequences reported in other bony fish, and it shares identical structural features. In addition, we analysed the fundamental property of CaBAFF protein usiong biological software. This study provided the basis for further investigation of its biological fuction in Carassius auratus and its relationship with imumune.

Related Dissertations

  1. Cloning and Expression Analysis of GPx, GST and SAHH Genes in Chlamydomonas Sp. ICE-L from Antarctica,Q943.2
  2. Research on Influnce of Silicon and Sulfur on Arsenic Uptake and Accumulation of Rice (Oryza Sativa L.),S511
  3. Effects of Dietary Oil Sources on Growth Perfomance, Lipid Deposition and Blood Biochemical Index of Carassius Auratus Gibelio,S963.1
  4. Cloning Andanalysis of NAC Transcription Factor Gene in Tobacco(Nicotiana Tabacum L.),Q943.2
  5. Cloning and Molecular Analysis of Lovastatin Biosynthesis Related Genes from Monascus Purpureus,TQ927
  6. Evaluation on Forage Quality and Biomass Energy Characters of Inbred Vegetative Lines of Napier Grass,S543.9
  7. Cloning and Function Analysis of P-ATPases Genes in Cotton and Tomato,S562
  8. Oshima Ye Lu Ju CcSOS1 Cloning and Expression Analysis,S682.11
  9. Silkworm circadian clock gene cloning and functional characterization Cry,S881.2
  10. The Characteristic of the Whole Hepatitis B Virus Gene, Construction of HBV Gene Eukaryotic Cell Expression Vector, and the Efficacy of Adefovir Plus Entecavir or Plus Lamivudine as Rescue Therapies in Chronic Hepatitis B Patients with Entecavir-resistant,R512.62
  11. Research on Cloning and Expression of the Bile Salt Hydrolase Gene from Lactobacillus Plantarum JPP2,Q78
  12. Identification of Resistance to Downy Mildew and Its Related Gene Cloning in Radish (Raphanus Sativus L.),S631.1
  13. The Transmembrane Fusion Protein of MnSOD Clone Expression and Function Study,Q78
  14. Günther Myostatin gene pathway gene cloning and expression,S917.4
  15. Fosmid library and PCR -based screening technologies spider silk protein gene package egg and Analysis,Q78
  16. Cloning and Expression Analysis of Hsp70 Gene from Freshwater Planarian Dugesia Japonica,Q78
  17. In some regions of porcine circovirus type 2 strains in Isolation, identification and analysis of whole-genome sequence,S852.65
  18. Sequence Analyzing and Prokaryotic Expression of ORF2 Gene in the Strain of Porcine Circovirus Type 2 Isolated from Lanzhou,S852.65
  19. Increase of the Yield of γ-decalactone by Genetically Modified Strain of Yarrowia Lipolytica,TS202.3
  20. Cloning Gonald-inhibiting Hormone Gene in Eriocheir Sinensis,S917.4

CLC: > Agricultural Sciences > Aquaculture, fisheries > Aquatic basic science > Aquatic Biology > Aquatic Zoology
© 2012 www.DissertationTopic.Net  Mobile