02221     2200205   4500001002100000005001500021035002000036245030000056700001900356700011000375250003800485260000900523300003200532084001500564082001500579008003900594650009800633520126900731990001502000INLIS00000000001769620211004094544  a0010-0721002228  acDNA library construction and isolation of genes for candidate vaccine antigens from Chrysomya bezziana (the Old World Screwworm fly). /cVuocolo,Tony,Supriyanti, Florentina; Muharsini, Sri;[Research Institute for Veterinary Science Jalan R.E. Martadinata 30, P.O. Box 151 Bogor 16114 Indonesia]  aMuharsini, Sri  a[Research Institute for Veterinary Science Jalan R.E. Martadinata 30, P.O. Box 151 Bogor 16114 Indonesia]  aJurnal Ilmu Ternak dan Veteriner.  c2000  aVol.5 (3): p.160-169. 2000.  aARTVET2042  aARTVET2042211004                |          | |   4acDNA library, peritrophin, peritrophic membrane, Chrysomya bezziana, Lucilia cuprina, vaccine  aThe construction and use of cDNA libraries for the isolation of genes encoding candidate antigens for use in a recombinant
vaccine against Chrysomya bezziana is described. RNA was isolated and mRNA purified from first and third instar larvae of 
Chrysomya bezziana and used in the synthesis of two cDNA libraries in the bacteriophage vector ? ZAP express
. These
libraries were screened using Digoxigenin-labeled DNA probes obtained from two independent approaches. First, a homolog
approach used probes designed from previously characterized peritrophic membrane genes identified from the related myiasis 
fly, Lucilia cuprina. Secondly, a de novo approach used amino-terminal and internal peptide sequence information derived from
purified Chrysomya bezziana peritrophic membrane proteins to generate DNA probes. Three peritrophic membrane genes were
identified and characterized. Chrysomya bezziana peritrophin-48 was identified using the homolog approach and, Chrysomya
bezziana peritrophin-15 and Chrysomya bezziana peritrophin-42 were identified using the de novo approach. The identification
of these genes as encoding candidate antigens against Chrysomya bezziana has allowed the production of recombinant proteins
for use in vaccination trials.  aARTVET2042