Beta-synuclein protein from Xenopus laevis: overexpression in Escherichia coli of the GST-tagged protein and production of polyclonal antibodies.
This report presents a procedure to obtain and purify recombinant beta-synuclein from Xenopus laevis expressed in Escherichia coli as a glutathione-S-transferase fusion protein. After identification by mass spectrometry, the protein was then used to raise anti-X. laevis beta-synuclein polyclonal antibodies, which were suitable to detect the presence of beta-synuclein in X. laevis brain by Western blot. This is the first report of a positive identification of beta-synuclein in an amphibian at the protein level.
Yuan
Zhu
Z
State Key Laboratory of Biotherapy, West China Hospital, West China Medical School and College of Life Science, Sichuan University, Chengdu, Sichuan, 610041, China.
Zhao
Xinyu
X
Yan
Fei
F
Zhao
Jian
J
Liu
Huanyi
H
Xiong
Shaoquan
S
Li
Jiong
J
Chen
Lijuan
L
Wei
Yuquan
Y
D000595:Amino Acid Sequence; D000818:Animals; D000917:Antibody Formation; D001921:Brain / Q000378:metabolism; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D008969:Molecular Sequence Data; D011817:Rabbits; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology; D021241:Spectrometry, Mass, Electrospray Ionization; D029867:Xenopus Proteins / Q000235:genetics* / Q000276:immunology* / Q000378:metabolism; D014982:Xenopus laevis / Q000235:genetics* / Q000276:immunology* / Q000378:metabolism; D051846:beta-Synuclein / Q000235:genetics* / Q000276:immunology* / Q000378:metabolism
D011993:Recombinant Fusion Proteins; D029867:Xenopus Proteins; D051846:beta-Synuclein