GTMB 7 - Gene Therapy & Molecular Biology

More documents

Recommendations

Info

Almazán et al: Expressed sequence tags in Ixodes scapularisTable 3. Summary of results of DNA vaccination and challenge with I. scapularis larvae in the mouse model of tickinfestations.EST cDNA clone Predicted protein Inhibition of tick infestation4D8Identity to H. sapiens hypothetical proteinFLJ10342 with unknown functionI (%)Inhibition of moltingM (%)40 a 7 a4F8 Nucleotidase 50 a 17 a1C10 b Hsp70 17 a 0 a3F6 b4D64E6Hsp60Identity to D. melanogaster CG10395 cDNAwith unknown functionIdentity to D. melanogaster CG13597 cDNAwith unknown function61 1120 ND3E1 Beta-adaptin 27 54G11 Chloride channel 38 3017 clones b Ribosomal proteins 15 a 0 a2C12 Beta-amyloid precursor protein (APP) -8 c ND4F1 Block of proliferation Bop1 -39 c ND3E10 Mannose binding lectin -48 a, c ND3C12 b RNA polymerase III -104 a, c ND2F9 bIdentity to A. variegatum AvGI TC255 &Homo sapiens hypothetical proteinFLJ12475 with unknown functions1A9, 1B2, 4A4 b ATPase -57 a, c NDa Data reported by Almazán et al. (2003). For all other experiments, mice were immunized with cDNA-containing expression plasmidDNA as described above. I and M were calculated as described in Materials and Methods section. ND, not determined.b Pooled together for vaccination experiments by ELI (Almazán et al, 2003) (1C10 and 3F6, cDNA pool “Heat shock”; 3C12 and 2F9,cDNA pool “Secreted protein”; ribosomal clones, cDNA pool “Ribosomal”; 1A9, 1B2 and 4A4, cDNA pool “ATPase”).c Resulted in enhanced tick feeding after mouse vaccination and tick challenge.The cDNA in clone 4G11 was identical to a chloridechannel but it contained only a partial sequence (Figure2A). This sequence protected against tick infestations andinhibited larval molting (Table 3). Chloride channels havebeen found in living organisms from bacteria to mammals,with some amino acid positions being conserved in allsequences (Figure 2A). As expected, phylogeneticanalysis of chloride channel sequences demonstrated thatthe I. scapularis sequence comprised a sister group toother insect sequences that have been reported (Figure2B).Vaccination with ribosomal sequences had someinhibitory effect on tick infestations but did not affectmolting (Table 3). The pool of ribosomal cDNAs includedEST sequences coding for cellular and mitochondrialribosomal proteins and translation factors (Table 4), andthese genes are highly conserved across species. However,proteins encoded by I. scapularis ESTs were 43% to 95%identical to arachnida or insect sequences and 36% to 85%identical to mouse sequences (Table 4). The cDNA inclone 2C12 that was found to be identical to the betaamyloidprecursor protein (APP) contained a fragmentencoding for the C-terminal of the protein (Figure 3),suggesting that it contains a partial cDNA with a long(1,400 bp) 3’ UTR. Nonetheless, the C-terminal sequenceof the I. scapularis APP contained regions of amino acidsidentical to fly and mosquito sequences (Figure 3).Vaccination with this cDNA resulted in 8% enhancementof larval feeding (Table 3). Vaccination with cDNA clone4F1 resulted in enhanced larval feeding (Table 3). Thecomplete sequence of clone 4F1 cDNA was determinedand contained an insert of 2,475 bp with 30 bp and 66 bpof 5’ and 3’ UTR, respectively and a poly-A tail of 114bases.An open reading frame of 2,265 bp encoded for aprotein of 754 amino acids that was identical to mouseblock of proliferation (Bop 1) (Figure 4).Similar proteins have been identified in otherorganisms including Drosophila melanogaster, Anophelesgambiae and humans (Figure 4), suggesting that thisprotein has been highly conserved during evolution. Theclone 3E10 had a pronounced stimulatory effect on larvalfeeding (Table 3). This clone was completely sequencedand contained an insert of 1,848 bp with 50 bp and 279 bpof 5’ and 3’ UTR, respectively and a short poly-A tail of24 bases. An open reading frame of 1,494 bp encoded fora protein of 497 amino acids that was identical tomannose-binding lectins found in many eukaryotes(Figure 5). A similar sequence was described in A.variegatum ESTs, which clustered together with the I.scapularis sequence (Figure 5).48
Gene Therapy and Molecular Biology Vol 7, page 49ABcgATGCAGGCGATGACGGGCTTTGCGGTGCAGTTCAACAAAAACAGTTTCGGGCTGACTCCAGCTCAGCCGCTGCAGTTGCAGATTCCCCTGCAGCCCAACTTCCCAGCTGATGCGAGCTTGCAGCTGGGAACCAACGGTCCCGTGCAGAAGATGGACCCCCTCACCAACCTTCAGGTGGCCATCAAGAACAATGTGGACGTGTTCTACTTCAGCTGCCTGGTGCCCATGCACGTGCTGAGCACGGAGGACGGCCTGATGGACAAGCGGGTGTTCCTGGCCACCTGGAAAGACATCCCCGCCCAAAACGAGGTCCAGTACACCCTCGACAACGTCAACCTCACTGCAGACCAAGTTTCCCAGAAGCTGCAGAACAACAACATTTTCACGATAGCCAAGAGGAACGTGGACGGCCAGGACATGCTGTACCAGTCCCTGAAGCTCACCAACGGCATTTGGGTGTTGGCGGAGCTCAAGATACAGCCCGGCAATCCAAGGATCACGTTGTCTTTGAAGACAAGAGCACCTGAAGTGGCAGCAGGTGTACAACAAACTTACGAACTCATTCTACACAGCTGAggctgctgtgaatgaaactcttctcccacccccttcttttgatggcagtcaatgtctcgtttcattttcttgttttcttttgcggcgtgctacggaacaaggtcctacattcccaagttatatggtgttgtcgcgtagggggcagagtgccgctgagcccgcgacagccttgtttctgaggagagccgaacgcaccacttcgaaaaagaaaaagtgaaaacggaaaaatgaaaaattttccagttgcttcaaattaacattcctcgtagtcagtctgtggccgttgagtttggtgtaaagaagaaaaaggtgtctcttttagtgaaaatggttgctttttattggtatcccctatcacaccgagcacgaacataagaaatcctgacaaggattctcctttagttgtattatggtggctggagcacacgaggcacctgttgccaattcgacccagcaaatgcccaattctcaagatttgagttcattgaggttgttttgctcctccccccccaccccccaactttgtcgttggattgtctaacagtgtaaatgggcgacgactcgttattctttttttcttcattctttctttttgttgtcacgcgccccgggggacgcgacacaacttatgtgcataattgattttcacaggctgcgacgcagtctgtaaaagaaggggaagtgaaactctgctccgccgctgctagtgtcatcacgggacgaccatcgcgttttctctgactatttaaacaaaactgcatagcttagggggcagtctgtgcaaagtggaacaaccaaactgagccctgccctttcggtgtgtgtacaagcatctctgtgtaacatgaactactttacatgaactacattgcatgaacgggagaagtttagttgtttttttgttttttttttcaggtgactatgtcaacagattagaaccattttttggaacggctggaaagataaccgctcattttgtttctactaaaagactacgaaaagtgttgactttttgcatcggtttggcaacgtttgtttggcatgcatgtagttgagcgtaatggtatcacccctcgtaaacaataacagtgcaatggagcagtactgtagtgtccattaaagagcgagagtttggttaaaggttgttaattgaggtccgtgttatcctttgagtaggagagcggcactttttgcaaatagcgctgctgggggcgtcatatctgccctccaaaacatgcacattttaagtgtgaattgttgcggcggcttgtacaagtatgtgtgttatgtgtagaaaaagaactcttaattaaaatatttgtggccaaaacgtcaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaM. musculus (747) LQHMTDFAIQFNKNSFGVIPSTPLAIHTPLMPNQSIDVSLPLNTLGPVMKD. melanogaster (731) MQPMTNFAIQLNKNSFGLVPASPMQ-AAPLPPNQSIEVSMALGTNGPIQRH. sapiens (68) LQHMTDFAIQFNKNSFGVIPSTPLAIHTPLMPNQSIDVSLPLNTLGPVMKI. scapularis (1) MQAMTGFAVQFNKNSFGLTPAQPLQLQIPLQPNFPADASLQLGTNGPVQKConsensus (748) LQHMTDFAIQFNKNSFGLIPATPLQIHTPLMPNQSIDVSLPLNTNGPVQKM. musculus (797) MEPLNNLQVAVKNNIDVFYFSCLIPLNVLFVEDGKMERQVFLATWKDIPND. melanogaster (780) MEPLNNLQVAVKNNIDIFYFACLVHGNVLFAEDGQLDKRVFLNTWKEIPAH. sapiens (118) MEPLNNLQVAVKNNIDVFYFSCLIPLNVLFVEDGKMERQVFLATWKDIPNI. scapularis (51) MDPLTNLQVAIKNNVDVFYFSCLVPMHVLSTEDGLMDKRVFLATWKDIPAConsensus (798) MEPLNNLQVAVKNNIDVFYFSCLIPLNVLFVEDGKMDKRVFLATWKDIPNM. musculus (847) ENELQFQIKECHLNADTVSSKLQNNNVYTIAKRNVEGQDMLYQSLKLTNGD. melanogaster (830) ANELQYTLSGVIGTTDGIASKMTTNNIFTIAKRNVEGQDMLYQSLKLTNNH. sapiens (168) ENELQFQIKECHLNADTVSSKLQNNNVYTIAKRNVEGQDMLYQSLKLTNGI. scapularis (101) QNEVQYTLDNVNLTADQVSQKLQNNNIFTIAKRNVDGQDMLYQSLKLTNGConsensus (848) ENELQFTIKEVHLTADTVSSKLQNNNIFTIAKRNVEGQDMLYQSLKLTNGM. musculus (897) IWILAELRIQPGNPNYTLSLKCRAPEVSQYIYQVYDSILKN-D. melanogaster (880) IWVLLELKLQPGNPEATLSLKSRSVEVANIIFAAYEAIIRSPH. sapiens (218) IWILAELRIQPGNPNYTLSLKCRAPEVSQYIYQVYDSILKN-I. scapularis (151) IWVLAELKIQPGNPRITLSLKTRAPEVAAGVQQTYELILHS-Consensus (898) IWILAELKIQPGNPNYTLSLKCRAPEVAQYIYQVYDSILKSFigure 1. Analysis of clone 3E1 identical to beta-adaptin. (A) Nucleotide sequence of complete cDNA. Non-coding sequence is shownin lower case letters and coding sequence is shown in capital letters with translation initiation and termination codons in bold letters. (B)Alignment of M. musculus (GenBank accession number XP_109938), D. melanogaster (CAA53509) and Homo sapiens (AAA35583)protein sequences and the translation product of clone 3E1 identified as I. scapularis beta-adaptin appendage (AY296113). Proteinsequences are shown in the single letter amino acid code. Identical amino acids are shown in red and amino acids conserved in 3 of 4sequences are shown in blue.AE. coli (4) DTPSLETPQAARLRRRQLIRQLLERDKTPLAILFMAAVVGTLVGLAA-VAO. mossambicus (98) DLKEGVCLSALWFNH--------EQ----------CCWTSNETTFAERDKX. laevis (146) DLKEGICLPWFWFNH--------EQ----------CCWQSNNVTFEDRNNI. scapularis (1) DLKEGICPQAFWLNK--------EQ----------CCWASNDTFFKG-DDC. elegans (141) DLKTGVCADRFWLDH--------EH----------CCWSSNDTFYKD-DDD. melanogaster (223) DLKHGICPPAFWFNR--------EQ----------CCYPAKQSVFEE-GNL. major (114) AFRSGICANFFWLGR-------------------------N-MCCVDCREA. gambiae (272) DLKFGICPQAFWLNR--------EQ----------CCWSSNETSFDS-GN49
Page 7 and 8:
Instructions to authors:Gene Therap
Page 9:
Please submit an electronic version
Page 12: 103-111 ResearchArticle113-133 Revi
Page 17 and 18: Gene Therapy and Molecular Biology
Page 61: Gene Therapy and Molecular Biology
Page 77: Gene Therapy and Molecular Biology
Page 80 and 81: Epperly et al: Late injection of Mn
Page 82 and 83: Epperly et al: Late injection of Mn
Page 84 and 85: Goldberg-Cohen et al: Regulation of
Page 90 and 91: Gascón-Irún et al: Gene therapy a
Page 106 and 107: Suzuki et al: Regulation of the Sp/
Page 112 and 113:
Suzuki et al: Regulation of the Sp/
Page 114 and 115:
Li et al: MET amplification in live
Page 116 and 117:
Li et al: MET amplification in live
Page 118 and 119:
Chavakis et al: Leukocyte adhesion
Page 120 and 121:
Page 122 and 123:
Page 124 and 125:
Page 126 and 127:
Page 128 and 129:
Sanlioglu et al: Adenovirus mediate
Page 130 and 131:
Page 132 and 133:
Page 134 and 135:
Page 136 and 137:
Page 138 and 139:
Page 140 and 141:
Page 142 and 143:
Page 144 and 145:
Page 146 and 147:
Page 148 and 149:
Page 150 and 151:
George et al: Gene therapy for vasc
Page 152 and 153:
Page 154 and 155:
Page 156 and 157:
Page 158 and 159:
Page 160 and 161:
Page 162 and 163:
Page 164 and 165:
Page 166 and 167:
Page 168 and 169:
Zhang et al: Angiogenic Gene Therap
Page 170 and 171:
Page 172 and 173:
Page 174 and 175:
Page 176 and 177:
Page 178 and 179:
Page 180 and 181:
Page 182 and 183:
Xu et al: G-CSF receptor-mediated S
Page 184 and 185:
Page 186 and 187:
Page 188 and 189:
Burek et al: Calcium induced cell d
Page 190 and 191:
Page 192 and 193:
Page 194 and 195:
Page 196 and 197:
David et al: Current status and fut
Page 198 and 199:
Page 200 and 201:
Page 202 and 203:
Page 204 and 205:
Page 206 and 207:
Page 208 and 209:
Page 210 and 211:
Page 212 and 213:
Page 214 and 215:
Page 216 and 217:
Page 218 and 219:
Page 220 and 221:
Page 222 and 223:
Page 224 and 225:
Page 226 and 227:
Stoll et al: The role of EBV and ge
Page 228 and 229:
Page 230 and 231:
Page 232 and 233:
Page 234 and 235:
Page 236 and 237:
Maruyama et al: Kidney-targeted pla
Page 238 and 239:
Page 240 and 241:
Page 242 and 243:
Page 244 and 245:
Kren et al: Hepatocyte-targeted del
Page 246 and 247:
Page 248 and 249:
Page 250 and 251:
Page 252 and 253:
Page 254 and 255:
Zeng: PRL-3 as a target for cancer
Page 256 and 257:
Page 258 and 259:
Page 260 and 261:
Latchman: Protective effect of heat
Page 262 and 263:
Page 264 and 265:
Page 266 and 267:
Page 268 and 269:
Page 270 and 271:
Cai et al: Lung cancer gene therapy
Page 272 and 273:
Page 274 and 275:
Page 276 and 277:
Page 278 and 279:
Page 280:
Page 283 and 284:
Gene Therapy and Molecular Biology
Page 285 and 286:
Page 287 and 288:
Page 289 and 290:
Page 291 and 292:
Page 293 and 294:
Page 295 and 296:
Page 297 and 298:
Page 299 and 300:
Page 301 and 302:
Page 303 and 304:
Page 305 and 306:
Page 307 and 308:
Page 309:
show all

GTMB 7 - Gene Therapy & Molecular Biology

Create successful ePaper yourself

Delete template?

Save as template?