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Prokaryotic Expression， Purification and Polyclonal Antibody Preparation of Chlamydomonas reinhardtii IFT22 Protein
Xue Bin1,2,3,4, Wang Gaofei1, Pan Qiang1, Li Shuting1, Wang Hao1, Li Xiaojin1, Fan Zhenchuan1,2,3,4*
1College of Food Engineering and Biotechnology, Tianjin University of Science & Technology, Tianjin 300457, China; 2Institute of Health Biotechnology, Tianjin University of Science & Technology, Tianjin 300457, China; 3International Collaborative Research Center for Health Biotechnology, Tianjin 300457, China; 4China International Science and Technology Cooperation Base for Health Biotechnology, Tianjin University of Science & Technology, Tianjin 300457, China
Abstract: Intraflagellar transport protein IFT22 is an important component of the IFT-B， but its function is controversial. It is indispensable to create a specific anti-IFT22 for its function research. Here， bacterial-expressed N-terminal His-tagged full length IFT22 recombinant protein was used as the antigen to immunize the New Zealand white rabbits. After the third immunization， the titer was determined to be 64 000 by an indirect ELISA. The antiserum was purified by Protein A and antigen， and the Western blot test showed only one band appeared when antigen was whole cell of Chlamydomonas reinhardtii CC125. The result showed the anti-IFT22 had a great specificify， which was an excellent material of studying function of IFT22.
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