Dissertation > Excellent graduate degree dissertation topics show

Cloning and Expression of Antimicrobial Peptides of Cathelicidin in Bovine Marrow

Author: XuPeng
Tutor: ZhangYong
School: Gansu Agricultural University
Course: Clinical Veterinary Medicine
Keywords: bovine mastitis Cathelicidins antimicrobial peptides BMAP-28 Indolicidin gene cloning prokaryotic expression antimicrobial activity
CLC: S858.23
Type: Master's thesis
Year: 2010
Downloads: 46
Quote: 0
Read: Download Dissertation

Abstract


Bovine Mastitis is one of the three diseases which can cause great loss in dairy industry and it is difficult for prevention and cure to the disease because long-term antibiotic therapy leads to the formation of resistants. Cathelicidin antibacterial peptides have very broad antimicrobial spectrum, which play an important role in innate immunity for mammals. In recent years, with the development of molecular biology, new peptide drugs have been developed, which hasl provide a new way for prevention and cure of mastitis.In this study, calf bone marrow was used for the material and total RNA was extracted. Indolicidin and BMAP-28 gene fragments were amplified by using RT-PCR and were cloned into pMD-18 vector. After identification by PCR, restriction enzyme digestion and sequencing analysis, the recombinant plasmid pMD18-T-Indolicidin and pMD18-T-BMAP28 were confirmed. BMAP-28 gene signal peptide was amplified by using recombinant plasmid pMD18-T-BMAP28 as a template, connected to the original expression vector pET-28a, and transformed into host strain Rosetta induced by IPTG. SDS-PAGE and Western-blot were used to detect expression product and protein was recycled. The antimicrobial activity of the target protein was tested by the major mastitis pathogens separated from a certain farm and standards strain purchased.The results showed that:1. There was one base difference was in Indolicidin gene between cloned and published. However, the amino acid sequences were not affected. There was one base difference between obtained the BMAP-28 gene sequences and published corresponding sequences,which led to amino acid substitutions.2. After induction by IPTG, Rosetta bacteria containing pET28a-BMAP28 recombinant plasmid expressed fusion protein of the molecular weight of about 19.3kda, which was consistent with the expected size.3. Isolated pathogens from Mastitis in dairy cows which mainly concluded Escherichia coli, Staphylococcus aureus and Streptococcus, and purchased standard strain were used for antimicrobial testing of recombinant BMAP-28 expressed protein. The expressed protein had a significant inhibition to resistance E.coli, resistance Streptococcus, standard E.coli and standard S.aureus, but a weak inhibitory effect on resistance S.aureus.

Related Dissertations

  1. Expression of D-AtCGS in E. Coli and Preparation of Polyclonal Antibody Against D-AtCGS,Q943.2
  2. Cloning and Expression of CHS and CHI Genes and Their Regulation on the Accumulation of Flavonoids in ’Cara Cara’ Navel Orange (Citrus Sinensis Osbeck) and ’Guoqing NO.4’ Satsuma Mandarin (Citrus Unshiu Marcow),S666.4
  3. Cloning and Expression Analysis of GPx, GST and SAHH Genes in Chlamydomonas Sp. ICE-L from Antarctica,Q943.2
  4. cDNA Cloning, Expression of vp5 and vp7 Genes and Subcecullar Localization of VP5 and VP7 Proteins in Grass Carp Reovirus,S941.41
  5. Cloning, Expression of vp6 and ns38 Genes and Immunogenicity of VP6 and NS38 in Grass Carp Reovirus,S941.41
  6. Cloning and Expression of Growth Hormone Gene in Cobia (Rachycentron Canadus),Q786
  7. Functional Analysis of Proteins Encoded by RNA2 of Wheat Yellow Mosaic Virus,S435.121
  8. Comparative Study on Reproductive Biological Characteristics of Helicoverpa Armigera and Helicoverpa Assulta (Lepidoptera:Nuctuidae),S433
  9. Cloning and Expression Analysis of Scavenger Receptor Class B Type Ⅰ and Antifreeze Proteinstype Ⅱ Genes in Lutjanus Sanguineus,S917.4
  10. Identification of B.melitensis 16M Protein Immunodominant Antigens,S852.61
  11. Molecular Cloning, Mrna Expression and Rnai of Nadph-Cytochrome P450 Reductase Gene in Helicoverpa Armigera (Hǘbner),S435.622.3
  12. Isolation and Characterization of Carbendazim-Degrading Strains, Cloning and Expression of the mheI Gene,X172
  13. Isolation and Identification of Infectious Bronchitis Virus and Sequence Analysis of Its S1 Gene and N Gene,S852.65
  14. Isolationand Identification of Porcine Parvovirus and Parts of Its Biological Characteristics,S852.65
  15. Isolation, Identification and Degrading Characteristic of a Cyhalofop-Butyl-Degrading Bacteria, and Cloning, Expression of a Cyhalofop-Butyl-Hydrolyzing Gene chbH,X172
  16. Cloning and Expression of Organic Solvent Tolerant Lipase Gene from Staphylococcus Saprophyticus M36,Q78
  17. Prokaryotic Expression and Purification of Human Beta-Defensin-9,Q78
  18. Molecular Cloning, Prokaryotic Expression and Characterization of a Novel Trypsin Inhibitor in the Frog, Lepidobatrachus Laevis,Q78
  19. Biochemical Characterization and Molecular Modification of a Novel Marine Mud-derived Pyrethroid Hydrolase,X172
  20. Cloning and Functional Analysis of Lytic Enzyme Production Related Gene Ctp in Lysobacter Enzymogenes Strain OH11,TQ925
  21. Screening of Bacterial Strains with High α-Galactosidase Activity and Its Gene Cloning, Expression, Purification and Characterization,TQ925

CLC: > Agricultural Sciences > Livestock, animal medicine,hunting,silkworm,bee > Animal Medicine ( Veterinary Medicine) > Livestock, poultry, wildlife diseases > Livestock > Cow
© 2012 www.DissertationTopic.Net  Mobile