水稻Qb-SNARE蛋白OsNPSN11多克隆抗体制备、鉴定与应用

doi:10.3724/SP.J.1005.2010.00961

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HEREDITAS ›› 2010, Vol. 32 ›› Issue (9): 961-965.doi: 10.3724/SP.J.1005.2010.00961

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Preparation, characterization and application of rice Qb-SNARE protein OsNPSN11 polyclonal antibody

BAO Yong-Mei, LIU Yong-Hui, XU Dong-Qing, HUANG Ji, WANG Zhou-Fei, WANG Jian-Fei, ZHANG Hong-Sheng

State Key Laboratory of Crop Genetics and Germplasm Enhancement, Nanjing Agricultural University, Nanjing 210095, China

Received:2009-12-30 Revised:2010-04-19 Online:2010-09-20 Published:2010-09-20
Contact: ZHANG Hong-Sheng E-mail:hszhang@njau.edu.cn

Abstract

Abstract: 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.

Key words: rice, Qb-SNARE protein, OsNPSN11, polyclonal antibody, expression analysis in transgenic plants

BAO Yong-Mei, LIU Yong-Hui, HU Dong-Qing, HUANG Ji, WANG Zhou-Fei, WANG Jian-Fei, ZHANG Gong-Sheng. Preparation, characterization and application of rice Qb-SNARE protein OsNPSN11 polyclonal antibody[J]. HEREDITAS, 2010, 32(9): 961-965.

References

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Preparation, characterization and application of rice Qb-SNARE protein OsNPSN11 polyclonal antibody

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