[Preparation, characterization and application of rice Qb-SNARE protein OsNPSN11 polyclonal antibody].
Membrane fusion in vesicle trafficking in the cells of eukaryotic organisms is mediated by soluble-N-ethyl- maleimide-sensitive fusion protein attachment protein receptor (SNARE) proteins. OsNPSN11 is a member of Qb-SNARE gene family isolate from rice. The cDNA of OsNPSN11 was subcloned into pET-30a and fusion to the 6 × His tag. Induced by 0.5 mmol/L IPTG for four hours, the recombinant protein was highly expressed in Escherichia coli, which was purified by Ni2+ -NTA His-bind resin affinity chromatography column to be used as an antigen to raise the antibody in New Zealand rabbits. Western blotting analysis showed that the antibody can specifically recognize the expressed antigen and the OsNPSN11 in plasma membrane protein from various rice tissues. This indicated that the antibody can be used for expres-sion analysis in transgenic rice.
Bao
Yong-Mei
YM
State Key Laboratory of Crop Genetics and Germplasm Enhancement, Nanjing Agricultural University, Nanjing 210095, China. yongmeibao@njau.edu.cn
Liu
Yong-Hui
YH
Xu
Dong-Qing
DQ
Huang
Ji
J
Wang
Zhou-Fei
ZF
Wang
Jian-Fei
JF
Zhang
Hong-Sheng
HS
D000818:Animals; D000906:Antibodies / Q000276:immunology*; D000917:Antibody Formation; D015153:Blotting, Western; D002352:Carrier Proteins / Q000737:chemistry; D004926:Escherichia coli / Q000235:genetics; D012275:Oryza / Q000737:chemistry*; D050766:Qb-SNARE Proteins / Q000276:immunology*; D011817:Rabbits; D011993:Recombinant Fusion Proteins / Q000276:immunology*; D050600:SNARE Proteins / Q000276:immunology
D000906:Antibodies; D002352:Carrier Proteins; D050766:Qb-SNARE Proteins; D011993:Recombinant Fusion Proteins; D050600:SNARE Proteins