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| Polyclonal Antibody Preparation and Application of C.reinhardtii LZTFL1 Protein |
| LIU Yan-Xia, FAN Zhen-Chuan |
| Institute of Health Biotechnology, International Collaborative Research Center for Health Biotechnology, College of Food Science and Biotechnology, Tianjin University of Science and Technology, Tianjin 300457, China |
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Abstract Leucine zipper transcription factor 1 (LZTFL1) is a protein associated with cilia signaling, and it is unclear about its function and mechanism in signal transduction. While Chlamydomonas reinhardtii as a model organism that studies the signal transmission of cilia, there is little research on the LZTFL1 protein, and no corresponding antibody has been detected. Based on this phenomenon, polyclonal antibody of LZTFL1 was prepared for subsequent study. The aim was to clone and express C. reinhardtii Lztfl1 gene in Escherichia coli (E. coli). The Lztfl1 gene of C.reinhardtii CC125 was amplified by RT-PCR, then was sub-cloned and heterologous expressed in E.coli BL21(DE3) using pET-28a (+) expression vector. Fusion protein 6×His-LZTFL1 was purified and immunized into New Zealand white rabbit so as to prepare polyclonal antibody. Finally, the sensitivity and specificity of the polyclonal antibody were detected through indirect ELISA and Western blot, respectively. The anti-LZTFL1 showed high sensitivity (1:512 000) and specificity. The above results indicate that Lztfl1 gene has been successfully cloned and expressed, and the rabbit anti-C. reinhardtii LZTFL1 polyclonal antibody has been prepared, which laid the foundation for the study of the structure and function of LZTFL1 protein in C. reinhardtii and its interaction in cilia signal transduction.
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Received: 08 May 2017
Published: 15 November 2017
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