The constituents of Microctonus sp. parasitoid venoms.
Purified RNA transcripts from venom glands dissected from the parasitoid wasp Microctonus hyperodae were copied, cloned and sequenced using traditional dideoxy sequencing methods. Using mass spectrometry analysis of the trypsinised PAGE gel protein bands we identified the RNA transcripts for the 3 most abundant proteins found in the venom and hence obtained their full protein sequence. Other abundant transcripts were also further sequenced. To reduce the effort required to obtain transcript information we dissected venom glands from a second parasitoid, Microctonus aethiopoides (Morocco biotype). The RNA transcripts were purified and reverse transcribed but instead of cloning the cDNA it was directly sequenced using Roche GS20 pyrosequencing. Results from a single GS20 sequencing run provided data similar to that obtained by the traditional methods used in analysing transcripts from M. hyperodae in a fraction of the time and cost. Comparing the transcripts between the two species showed that a similar range of genes are expressed with the putative orthologs of seven of the eight full length genes characterised from M. hyperodae being found in M. aethiopoides. Pyrosequencing should provide a valuable new method for rapidly sampling transcripts from a wide range of specialised insect tissues.
Crawford
A M
AM
AgResearch Invermay Agricultural Centre, Puddle Alley, Private Bag 50034, Mosgiel, New Zealand. allan.crawford@agresearch.co.nz
Brauning
R
R
Smolenski
G
G
Ferguson
C
C
Barton
D
D
Wheeler
T T
TT
McCulloch
A
A
D000595:Amino Acid Sequence; D000825:Animal Structures / Q000378:metabolism; D000818:Animals; D001483:Base Sequence; D018076:DNA, Complementary / Q000235:genetics; D004210:Dissection; D015723:Gene Library; D019476:Insect Proteins / Q000737:chemistry / Q000235:genetics; D008969:Molecular Sequence Data; D010271:Parasites / Q000737:chemistry*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D017422:Sequence Analysis, DNA; D014862:Wasp Venoms / Q000737:chemistry*; D014863:Wasps / Q000737:chemistry*
D018076:DNA, Complementary; D019476:Insect Proteins; D012333:RNA, Messenger; D014862:Wasp Venoms