[Analysis of insecticidal crystal proteins from Bacillus thuringiensis strain 4.0718 through two-dimensional gel electrophoresis and MALDI-TOF-mass spectrometry].
The present study analyses the insecticidal crystal proteins (ICPs) from Bacillus thuringiensis strain 4.0718 through two-dimensional electrophoresis and matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF-MS). By comparing and optimizing the composition of lysis solution, the volume of sample loading and the protocol for isoelectric focusing, a well-focused 2-DE map with high resolution and reproducibility was achieved for the first time. And after an tryptic enzymolisis and a test of part of protein spots by means of MALDI-TOF-MS, the peptides mass fingerprint (PMF) was obtained and, by referring to Swiss-Prot, Cry1Ac and Cry2Aa contained in Bt 4.0718 were identified, with their molecular weights 134160 Da and 71097 Da respectively.
Song
Sheng
S
Microbiology Key Laboratory, Hunan Normal University, Hunan Province, Changsha 410081, China. ecosystem2000@tom.com
Xia
Li-qiu
LQ
Huang
Jiang-li
JL
Sun
Yun-jun
YJ
Ding
Xue-zhi
XZ
D001413:Bacillus thuringiensis / Q000737:chemistry*; D000083722:Bacillus thuringiensis Toxins; D001426:Bacterial Proteins / Q000032:analysis* / Q000737:chemistry; D001427:Bacterial Toxins / Q000032:analysis* / Q000737:chemistry; D015180:Electrophoresis, Gel, Two-Dimensional; D004731:Endotoxins / Q000032:analysis* / Q000737:chemistry; D006460:Hemolysin Proteins; D008970:Molecular Weight; D020543:Proteome / Q000032:analysis; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
D000083722:Bacillus thuringiensis Toxins; D001426:Bacterial Proteins; D001427:Bacterial Toxins; D004731:Endotoxins; D006460:Hemolysin Proteins; D020543:Proteome; C030956:insecticidal crystal protein, Bacillus Thuringiensis