[An establishmented methods of researching silk gland of silkworm(Bombyx mori L.) with proteomics].
We separated proteins of the middle silk gland through high resolution two-dimensional polyacrylamide gel electrophesis, and identified the high expressional proteins using MALDI-TOF-MS and analyzed the PMF in protein database by GPMAW( General Protein/Mass Analysis for Windows) . The protein database was forecasted through silkworm genome database. More than 500 spots were obtained from each gel by silver stain and more than 100 protein spots were obtained from gel by Coomassie brilliant blue stain. Most of them were distributed in the area from 15kD to 90kD with pH 3.5-7. Among the 25 Coomassie brilliant blue stained spots identified by MALDI-TOF-MS, more than 60% have relatively strong signal. Comparing with the result of using Mascot, the method using PMF database of silkworm not only can identify some known proteins, but also can identify some unknown proteins that have been forecasted in silkworm genome database. Accordingly, we founded a complete set of method that fit for researching proteome of silkworm.
Liu
Hong-Li
HL
The Key Sericultural Laboratory of Agricultural Ministry, Southwest University, Chongqing 400716, China.
Xia
Qing-You
QY
Hou
Yong
Y
Zhao
Ping
P
Zou
Yong
Y
Guan
Jian
J
D000595:Amino Acid Sequence; D000818:Animals; D012831:Bombyx / Q000378:metabolism*; D030562:Databases, Protein; D015180:Electrophoresis, Gel, Two-Dimensional; D006863:Hydrogen-Ion Concentration; D019476:Insect Proteins / Q000032:analysis* / Q000737:chemistry; D008969:Molecular Sequence Data; D008970:Molecular Weight; D020543:Proteome / Q000032:analysis* / Q000737:chemistry; D040901:Proteomics / Q000379:methods*; D047011:Silk / Q000378:metabolism*; D012984:Software; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
D019476:Insect Proteins; D020543:Proteome; D047011:Silk