Molecular Characterization and Gene Expression Profiling ... - CUSAT

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Chapter 1 naturally occurring peptides serve as starting points for the development of new drugs. Several derivatives of AMPs have been through the pharmaceutical process, including human phase I–III studies. The use of human AMPs as drug is restricted so far by the still unknown biological function of these molecules and the high cost for the generation of sufficient amount (Bals, 2000). Most clinical trials proposed or underway involve topical therapy. Such treatments are likely to be effective and safe because the more toxic cationic peptides and lipopeptides, including gramicidin S and polymyxin B, are already in skin creams. Cationic peptides with excellent activity against a broad range of bacteria would be especially indicated where there is a risk from seriously resistant pathogens or where current treatments are ineffective. A more advanced form of topical treatment would be aerosol therapy into the lung, an organ especially prone to problems with resistant microorganisms. Aerosol treatment is already used successfully for the polycationic trisaccharides gentamicin and tobramycin and the polycationic lipopeptide colistin in treatment of P. aeruginosa lung infections in patients with cystic fibrosis. These antibiotics are usually given in liposomal formulation and show little toxicity and reasonable efficacy long term. Oral therapy may be possible for gastrointestinal infections; nisin is being developed through to clinical trial in Helicobacter pylori infection (Hancock, 1997). Various applications of AMP are given in Table 1.5. Table 1.5 Applications of AMP AMP Source Application alfAFP Plant (Alfalfa) Antifungal effect in transgenic potato Cecropin B Insect CEMA Synthetic (Cecropinmelittin) Antibacterial effect in transgenic rice, antiviral effect against important viral pathogens of fish Antifungal effect in transgenic tobacco, antibacterial effect against bacterial pathogen in fish Molecular Characterization and Gene Expression Profiling of Antimicrobial Peptides in Penaeid Shrimps 86
Lactoferricin Cow Mastitis control in goat LzP Hen (Lysozyme) Food preservative MSI-99 Synthetic (Magainin analog) Nisin Lactococcus lactis Food preservative Antifungal and antibacterial effect in transgenic tobacco, tomato, grapevine, banana Polyoxins Streptomyces Fungicide, insecticide Pn-AMP2 MSI-78 IB367 NEUPREX1 MX-226 / CPI-226 Plant (Morning glory) Synthetic (Magainin analog) Pig protegrin analogue Synthetic (Cationic peptide fragment) Bovine indolicidinbased MX594AN Indolicidin-based Polymyxin B & E Gramicidin S hLF1-11 Bacillus polymyxa Antifungal effect in transgenic tobacco Wound healing (Phase III clinical trial) Against polymicrobic infections in oral mucasitis Chapter 1 For endotoxin mediated complications of meningococcal disease (Phase III clinical trial) Prevention of contamination of central venous catheters (Phase III clinical trial) Against mild to moderate acne (Phase II clinical trial) Food preservative, against multi-drug resistant Pseudomonas, Topical application Bacillus brevis Food preservative, Topical application Human lactoferricin-based Daptomycin Streptomyces roseosporus Colomycin Bacillus polymyxa Systemic application Pexiganan Magainin analog MX-226 / CPI-226 Cationic peptide MX594AN Indolicidin-based For the prevention of infections in patients undergoing allogeneic stem cell transplantation FDA approved (2003) - for the treatment of skin infections caused by S. aureus For the treatment of diabetic foot ulcer infections (Phase III clinical trial) For the prevention of catheter-related infections (Phase III clinical trial) For the treatment of acne (Phase II clinical trial) Molecular Characterization and Gene Expression Profiling of Antimicrobial Peptides in Penaeid Shrimps 87
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Molecular Characterization and Gene
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Declaration I hereby do declare tha
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Acknowledgements This work was carr
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My Seniors, Dr. Annies Joseph, Dr.
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Contents Title Page Nos. 1 General
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in response to WSSV challenge 252-2
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α-2M Alpha-2-Macroglobulin ALF Ant
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ppA Prophenoloxidase-Activating Enz
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1.1 Introduction Chapter 1 Aquacult
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Chapter 1 the giant tiger shrimp (P
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Common names: Chapter 1 Giant tiger
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1.3.1. Diseases Chapter 1 Diseases
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Lymphoidal parvo-like virus (LPV) B
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Chapter 1 and protective occlusion
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Chapter 1 small brown masses in the
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1.4.6. Natural epidemics Chapter 1
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Chapter 1 can differentiate WSSV-in
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1.5.1.1. Probiotics Chapter 1 Alter
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Chapter 1 tried in P. monodon, resu
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Chapter 1 The first and essential i
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Chapter 1 1998). The innate immunit
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1.6.1 Haemocytes Chapter 1 The haem
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Chapter 1 In crustaceans, the sheet
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Chapter 1 Theopold et al., 2002). C
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Chapter 1 kill/remove the invading
Page 51 and 52: Chapter 1 forming complexes with a
Page 53 and 54: 1.7 Antimicrobial Peptides (AMPs) 1
Page 55 and 56: Chapter 1 the response is very fast
Page 57 and 58: Chapter 1 Hirsch, 1947). While they
Page 59 and 60: Chapter 1 AMPs to various degrees u
Page 61 and 62: 1.7.5 Biological activity Chapter 1
Page 63 and 64: Chapter 1 including the microbes as
Page 65 and 66: Tachyplesin I Chapter 1 Crustacea P
Page 67 and 68: Anionic and cationic peptides that
Page 69 and 70: Anionic and Cationic AMPs with cyst
Page 71 and 72: Chapter 1 Fig. 1.7. Structural clas
Page 73 and 74: Chapter 1 Group III: Linear peptide
Page 75 and 76: Chapter 1 through regular processes
Page 77 and 78: Chapter 1 ultimately signal to tran
Page 79 and 80: Chapter 1 divalent polyanionic cati
Page 81 and 82: Chapter 1 type of transmembrane por
Page 83 and 84: Chapter 1 other intracellular targe
Page 85 and 86: Chapter 1 Apidaecin, a short, proli
Page 87 and 88: Chapter 1 Hultmark, 1987; Gennaro a
Page 89 and 90: Chapter 1 quite opposite characteri
Page 91 and 92: Chapter 1 depletion of mitochondria
Page 93 and 94: Chapter 1 conformations adopted by
Page 95 and 96: Chapter 1 receptor may be a potenti
Page 97 and 98: Chapter 1 exposure to their targets
Page 99 and 100: Chapter 1 generating a rich spectru
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Page 105 and 106: Chapter 1 trial, in which peptide T
Page 107 and 108: Chapter 1 a too limited spectrum to
Page 109 and 110: Chapter 1 number of research groups
Page 111 and 112: CHAPTER-2 Molecular Characterizatio
Page 113 and 114: Chapter 2 interaction with negative
Page 115 and 116: Chapter 2 inhibit the endotoxin or
Page 117 and 118: Chapter 2 structure be modified to
Page 119 and 120: Chapter 2 and the region might be l
Page 121 and 122: Chapter 2 been made of the spectra
Page 123 and 124: Chapter 2 their discovery and initi
Page 125 and 126: Chapter 2 terminal pyroglutamic aci
Page 127 and 128: Chapter 2 One can assume that the p
Page 129 and 130: Chapter 2 by degranulation into the
Page 131 and 132: 2.2. Materials and Methods 2.2.1. E
Page 133 and 134: Chapter 2 washed by adding 1 ml 75
Page 135 and 136: Chapter 2 entry of recombinant DNA
Page 137 and 138: Chapter 2 and the deduced amino aci
Page 139 and 140: Chapter 2 Fenneropenaeus, Litopenae
Page 141 and 142: 2.3.1.3. Crustin-1 (GQ334395) Chapt
Page 143 and 144: 2.3.1.3.5. Phylogenetic analysis of
Page 145 and 146: Chapter 2 sequence also showed high
Page 147 and 148: Chapter 2 chinensis and F. indicus
Page 149 and 150: Chapter 2 monodon and Group III of
Page 151 and 152: Chapter 2 Tiger shrimp (P. monodon)
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Chapter 2 ALF-1 was a partial seque
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Chapter 2 As mentioned before, the
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Chapter 2 Munoz et al., 2004). The
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Table 2.1. Primers used for the stu
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Table 2.4. BLASTn analysis of ALF-2
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Table 2.8. BLASTn analysis of Crust
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Table 2.12. BLASTn analysis of Pena
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Fig.2.5. Agarose gel electrophoreto
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Chapter 2 Fig.2.9. Multiple alignme
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Chapter 2 Fig. 2.12 A bootstrapped
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Chapter 2 Fig. 2.15. Multiple align
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Chapter 2 Fig.2.18. A bootstrapped
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Chapter 2 Fig.2.20. Signal peptide
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Chapter 2 Fig.2.24 Multiple alignme
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Chapter 2 LITOPENAEUS SP. FARFANTEP
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Chapter 2 Fig. 2.28. Nucleotide and
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Chapter 2 LITOPENAEUS SP. FENNEROPE
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Chapter 2 Fig.2.43 A bootstrapped n
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Chapter 2 LITOPENAEUS SP. FENNEROPE
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Chapter 2 Fig.2.51 Signal peptide a
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Chapter 2 FENNEROPENAEUS SP. FARFAN
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CHAPTER-3 Molecular Characterizatio
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Chapter 3 certainly help us to unra
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Chapter 3 3.3.1.1. Anti-lipopolysac
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Chapter 3 was predicted out to be 1
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Chapter 3 random coiled structure t
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3.3.1.3.5. Phylogenetic analysis of
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Chapter 3 case has been reported in
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Chapter 3 Multiple alignments were
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Chapter 3 negative bacteria. In shr
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Chapter 3 indicated a random coiled
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Chapter 3 horseshoe crab big defens
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Table 3.4. BLASTn analysis of ALF-2
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Table.3.8. BLASTn analysis of Fi-pe
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Chapter 3 Fig.3.2. Nucleotide and a
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Chapter 3 Fig.3.5. Multiple alignme
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Chapter 3 Fig.3.8. Nucleotide and a
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Chapter 3 Fig.3.12. Multiple alignm
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Chapter 3 Fig.3.14. A bootstrapped
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SHRIMPS KIND CRAB LOBSTERS CRABS Ch
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Chapter 3 Fig. 3.28. Multiple align
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PENAEIDIN-2 Chapter 3 PENAEIDIN- 3
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Fig.3.31. Nucleotide and amino acid
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Chapter 3 Fig.3.37 Nucleotide and a
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4.1. Introduction Chapter 4 The aqu
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Chapter 4 appearance and have major
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Chapter 4 the tissues has recently
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Chapter 4 with the ability of shrim
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Chapter 4 with a low dose of WSSV b
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Chapter 4 molecules they are likely
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Chapter 4 for understanding gene ex
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Chapter 4 4.3.2. Expression profile
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Expression profile of endonuclease
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Chapter 4 bacterial and fungal infe
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Chapter 4 measuring TSV and YHV loa
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Chapter 4 Of particular interest he
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Chapter 4 large distribution and ab
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Chapter 4 WSSV genes viz. endonucle
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Table 4.2. Primers used for the stu
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(A) (B) B 28 1 2 3 4 5 6 7 8 9 10 C
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(A) (B) B 28 1 2 3 4 5 6 7 8 9 10 C
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(A) (B) B 28 1 2 3 4 5 6 7 8 9 10 C
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(A) (B) B 28 1 2 3 4 5 6 7 8 9 10 C
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(A) (B) B 28 1 2 3 4 5 6 7 8 9 10 F
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(A) (B) B 28 1 2 3 4 5 6 7 8 9 10 C
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(A) (B) B 28 1 2 3 4 5 6 7 8 9 10 C
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(A) (B) B 28 1 2 3 4 5 6 7 8 9 10 C
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CHAPTER-5 Expression Profile of Ant
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Chapter 5 1994). The emergence of b
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Chapter 5 (Bovill et al., 2001). In
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5.2.2. Test diets Chapter 5 Four ex
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5.2.2.4. Preparation of Glucan inco
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Chapter 5 genes (latency related ge
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Chapter 5 On WSSV challenge, crusti
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Chapter 5 5.3.4. Expression profile
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5.3.4.5. Expression profile of pena
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Chapter 5 must firstly rest on a so
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Chapter 5 For the present study two
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Chapter 5 ameobocytes of two marine
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Chapter 5 variation in expression o
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Chapter 5 It has been previously sh
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Chapter 5 manifested in terms of gr
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(A) (B) (C) C CHY CSY CHG CSG C CHY
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(A) (B) -VE CTRL +VE CTRL CHG CSG C
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(A) PRE-CHALLENGE (B) POST-CHALLENG
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(A) PRE-CHALLENGE (B) (C) (D) CONTR
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(C) (D) (A) PRE-CHALLENGE (B) CONTR
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(A) (B) G M HP HR I Chapter 5 Fig.
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Chapter 5 Fig. 5.26. Post challenge
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6.1. Introduction Chapter 6 Prophyl
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Chapter 6 production of inhibitory
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6.2. Materials and methods 6.2.1. E
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Chapter 6 tissue cDNA was performed
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Chapter 6 gene was supported by Bac
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Chapter 6 treated group of shrimps.
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Chapter 6 the gene was found to be
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Chapter 6 found to support minimum
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Chapter 6 is induced upon bacterial
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Chapter 6 all the target genes, exc
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Chapter 6 Detailed tissue-wise anal
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Chapter 6 Gills and intestine was f
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(A) (B) Fig. 6.1. Experimental diet
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(A) (B) (C) Fig. 6.3. Expression pr
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(A) (B) (C) C B M BM C B M BM Fig.
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(A) (B) (C) Fig. 6.7. Expression pr
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(A) (B) (C) Chapter 6 Fig. 6.9. Exp
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(A) (B) -VE CTRL +VE CTRL B M BM Ch
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(A) (C) (D) PRE-CHALLENGE POST-CHAL
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(A) (B) G M HP HR I Chapter 6 Fig.
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Chapter 6 Fig. 6.27. Post challenge
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7.1. Summary Chapter 7 Tropical shr
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Salient findings Chapter 7 � From
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Chapter 7 � Tissue-wise expressio
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Chapter 7 application in shrimp cul
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References Aboudy, Y., Mendelson, E
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References Anggraeni, M.S., Owens,
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References Bals, R., Wang, X., Zasl
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References derived from the N-termi
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References Breukink, E., de Kruijff
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References Burgents, J.E., Burnett,
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References 1,3-β-D-glucan-binding
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References Chen, J.Y., Chuang, H.,
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References Cole, A.M., Hong, T., Bo
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References Dathe, M., Wieprecht, T.
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References Diamond, G., Zasloff, M.
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References Faber, C., Stallmann, H.
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References Flint, S.J., Enquist, L.
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References Gennaro, R., Zanetti, M.
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References Gudmundsson, G.H., Lidho
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References Harwig, S.S., Swiderek,
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References Hirakura, Y., Kobayashi,
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References Huang, C.C., Song, Y.L.
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References Itami, T., Takahashi, Y.
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References Jiravanichpaisal, P. Whi
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References Kang, J.H., Shin, S.Y.,
Page 461 and 462:
References Kono, T., Savan, R., Sak
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References rich peptide derived fro
Page 465 and 466:
References Li, L., Wang, J.X., Zhao
Page 467 and 468:
References Liu, C.H., Cheng, W., Ku
Page 469 and 470:
References Lorenzon, S., de Guarrin
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References Marone, M., Mozzetti, S.
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References Medvinsky, A., Dzierzak,
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References Moriarty, D.J.W. 1996. M
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References antimicrobial peptide fr
Page 479 and 480:
References Otta, S.K., Karunasagar,
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References simplex virus has heptad
Page 483 and 484:
References infected shrimp. Thesis
Page 485 and 486:
References Litopenaeus vannamei cha
Page 487 and 488:
References Roth, B.L., Poot, M., Yu
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References Sathish, S., Musthaq, S.
Page 491 and 492:
References a proline-arginine-rich
Page 493 and 494:
References Smith, V.J., Fernandes,
Page 495 and 496:
References Song, Y.L., Cheng, W., W
Page 497 and 498:
References experimental system. In:
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References the black tiger shrimp P
Page 501 and 502:
References Tapay, L.M., Cesar, E.,
Page 503 and 504:
References Touhy, K.M., Probert, H.
Page 505 and 506:
References van Hulten, M.C., Reijns
Page 507 and 508:
References Vives, R.R., Imberty, A.
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References Wang, Y.C., Chang, P.S.,
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References Williams, M.J., Rodrigue
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References Wu, W., Zong, R., Xu, J.
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References Yodmuang, S., Tirasophon
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References Zhan, W.B., Wang, Y.H. 1
Page 519 and 520:
Penaeus monodon anti-lipopolysaccha
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Penaeus monodon crustin-like antimi
Page 523 and 524:
Penaeus monodon crustin-like antimi
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Penaeus monodon penaeidin-like anti
Page 527 and 528:
Penaeus monodon elongation factor m
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Fenneropenaeus indicus anti-lipopol
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Fenneropenaeus indicus penaeidin-li
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Fenneropenaeus indicus beta-actin m
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PUBLICATIONS
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Short sequence report Molecular cha
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Fig. 2. Multiple alignment of nucle
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220 The phylogenetic relationships
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Table 1 Rearing conditions and wate
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5 ′ cttgtggttgacaatggctccg3
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Expression of WSSV genes in the hae
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Fig. 4. Expression profile of crust
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S.P. Antony et al. / Immunobiology
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