Molecular Characterization and Gene Expression Profiling ... - CUSAT

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1.7.6.1 AMPs in marine invertebrates Chapter 1 The ocean covers 71% of the surface of the earth and contains approximately half of the total global biodiversity, with estimates ranging between 3 and 500 x 10 6 different species (Haug et al., 2002). Marine macrofauna alone comprise 0.5 to 10 x 10 6 species (de Vries and Hall, 1994). Therefore, the marine environment, especially marine invertebrates that rely solely on innate immune mechanisms for host defense, is a spectacular resource for the development of new antimicrobial compounds. Table 1.2 gives an overview of AMPs isolated and characterized from the marine invertebrates covering the phyla Porifera, Cnidaria, Annelida, Mollusca, Arthropoda, Echinodermata and Urochordata. Table 1.2. Major AMPs/AMP families isolated from marine invertebrates SOURCE ORGANISM AMP Phylum Group Species Porifera Sponges Discodermia kiiensis Discodermin Cnidaria Jelly Fish Aurelia aurita Aurelin Annelida Polychaetes Arenicola marina Arenicin Nereis diversicolor Hedistin Perinereis aibuhitensis Perinerin Mollusca Bivalves Mytilus edulis Defensins A and B Arthropoda Limulus Myticin Mytilin A and B Mytimicin Crassostrea virginica Defensin Mytilus galloprovincialis Myticin A and B Limulus polyphemus Defensins 1 and 2 Mytilin B, C, D, and G1 Polyphemusin Tachyplesin II Tachyplesus gigas Tachyplesin III Tachycitin Tachypleus tridentatus Big defensin Tachystatin Molecular Characterization and Gene Expression Profiling of Antimicrobial Peptides in Penaeid Shrimps 48
Tachyplesin I Chapter 1 Crustacea Pacifastacus leniusculus Astacidin, Pl crustin, ALF Carcinus maenas Carcinin, ALF Callinectes sapidus Callinectin, ALF Hyas araneus Hyasin, arasin Scylla paramanosian ALF, crustin Scylla serrata ALF, crustin Eriocheir sinensis ALF Panulirus argus Crustin, PET-15 Homarus americanus Crustin, Homarin Homarus gammarus Crustin Litopenaeus vannamei Penaeidin-2, -3, -4, crustin Litopenaeus setiferus Penaeidin-2, -3, -4, crustin, ALF Litopenaeus stylirostris Penaeidin-2, -3 Farfantepenaeus paulensis Penaeidin-2 Litopenaeus schmiitti Penaeidin-2, -4 Penaeus monodon Penaeidin-3, -5, crustin, Fenneropenaeus chinensis Echinodermata Sea Urchin Strongylocentrotus droebachiensis ALF Penaeidin-3, crustin, ALF Penaeus semisulcatus Penaeidin-3 Marsupenaeus japonicus Crustin, ALF Strongylocin Urochordata Tunicates Styela clava Styelin, Clavanin, Clavaspirin Halocynthia aurantium Dicynthaurin, Halocidin Halocynthia papillosa Halocyamine Halocynthia papillosa Papillosin Styela plicata Plicatamide Molecular Characterization and Gene Expression Profiling of Antimicrobial Peptides in Penaeid Shrimps 49
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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
Page 13 and 14: α-2M Alpha-2-Macroglobulin ALF Ant
Page 15 and 16: ppA Prophenoloxidase-Activating Enz
Page 17 and 18: 1.1 Introduction Chapter 1 Aquacult
Page 19 and 20: Chapter 1 the giant tiger shrimp (P
Page 21 and 22: Common names: Chapter 1 Giant tiger
Page 23 and 24: 1.3.1. Diseases Chapter 1 Diseases
Page 25 and 26: Lymphoidal parvo-like virus (LPV) B
Page 27 and 28: Chapter 1 and protective occlusion
Page 29 and 30: Chapter 1 small brown masses in the
Page 31 and 32: 1.4.6. Natural epidemics Chapter 1
Page 33 and 34: Chapter 1 can differentiate WSSV-in
Page 35 and 36: 1.5.1.1. Probiotics Chapter 1 Alter
Page 37 and 38: Chapter 1 tried in P. monodon, resu
Page 39 and 40: Chapter 1 The first and essential i
Page 41 and 42: Chapter 1 1998). The innate immunit
Page 43 and 44: 1.6.1 Haemocytes Chapter 1 The haem
Page 45 and 46: Chapter 1 In crustaceans, the sheet
Page 47 and 48: Chapter 1 Theopold et al., 2002). C
Page 49 and 50: 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: Chapter 1 including the microbes as
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
Page 101 and 102: Chapter 1 by enkelytin (Goumon et a
Page 103 and 104: Lactoferricin Cow Mastitis control
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
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Chapter 2 inhibit the endotoxin or
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Chapter 2 structure be modified to
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Chapter 2 and the region might be l
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Chapter 2 been made of the spectra
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Chapter 2 their discovery and initi
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Chapter 2 terminal pyroglutamic aci
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Chapter 2 One can assume that the p
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Chapter 2 by degranulation into the
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2.2. Materials and Methods 2.2.1. E
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Chapter 2 washed by adding 1 ml 75
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Chapter 2 entry of recombinant DNA
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Chapter 2 and the deduced amino aci
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Chapter 2 Fenneropenaeus, Litopenae
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2.3.1.3. Crustin-1 (GQ334395) Chapt
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2.3.1.3.5. Phylogenetic analysis of
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Chapter 2 sequence also showed high
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Chapter 2 chinensis and F. indicus
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Chapter 2 monodon and Group III of
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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.,
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References Kono, T., Savan, R., Sak
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References rich peptide derived fro
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References Li, L., Wang, J.X., Zhao
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References Liu, C.H., Cheng, W., Ku
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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
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References Otta, S.K., Karunasagar,
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References simplex virus has heptad
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References infected shrimp. Thesis
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References Litopenaeus vannamei cha
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References Roth, B.L., Poot, M., Yu
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References Sathish, S., Musthaq, S.
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References a proline-arginine-rich
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References Smith, V.J., Fernandes,
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References Song, Y.L., Cheng, W., W
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References experimental system. In:
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References the black tiger shrimp P
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References Tapay, L.M., Cesar, E.,
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References Touhy, K.M., Probert, H.
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References van Hulten, M.C., Reijns
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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
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Penaeus monodon anti-lipopolysaccha
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Penaeus monodon crustin-like antimi
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Penaeus monodon crustin-like antimi
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Penaeus monodon penaeidin-like anti
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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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