Abstract
In the studies of developmental gene expressions in a silkworm, it is necessary to attempt exhaustive protein analysis, proteomic analysis, with EST analysis. We started the silkworm proteomics using Daizo which has been studied EST. Middle silkgrand, posterior silkgrand, midgut, fat body and other tissues were collected from fifth-instar day-3 and 4 larva. Proteins were separated by a two-dimensional polyacrylamide gel electrophoresis and the each spot was digested by a trypsin after the alkylation. Peptides were analyzed by an on-line capillary HPLC-MS (LCQ Deca). The data obtained by MS analysis were applied to the program Mascot and Sequest for the identification of the proteins using the genome data from fruit fly and the translated data from silkworm EST. Many genes encoding protein were identified by MS analysis. Some of them were not hit to fruit fly genome data but did to silkworm EST. Additionally, artificial shift of reading frame in EST nucleotide sequences were found by MS analysis.
