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Issue:ISSN 1000-7083
          CN 51-1193/Q
Director:Sichuan Association for Science and Technology
Sponsored by:Sichuan Society of Zoologists; Chengdu Giant Panda Breeding Research Foundation; Sichuan Association of Wildlife Conservation; Sichuan University
Address:College of Life Sciences, Sichuan University, No.29, Wangjiang Road, Chengdu, Sichuan Province, 610064, China
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Your Position :Home->Past Journals Catalog->2011 Vol.30 No.4

Purification and Preparation of Anti-serum to Spider Dragline Silk Protein
Author of the article:WANG Chun-sheng, LUO Fang, ZHANG Zhi-ren, ZHAO Jian-ling, PIAO Shan-hua, AN Tie-zhu*
Author's Workplace:(College of Life Science, Northeast Forestry University, Harbin 150040,China)
Key Words:spider dragline silk protein; prokaryotic expression; protein purification; preparation of anti-serum
Abstract:In order to prepare anti-serum to spider dragline silk protein for in vitro detection, a recombinant prokaryotic expression plasmid 2S-pET-52b(+) was constructed and then expressed in BL21-pLysS cells for producing recombinant protein which was used as antigen after purification by the means of affinity choromatography. The results were showed as followed: recombinant plasmid 2S-pET-52b(+) was transformed into BL21-pLysS cells, and then selected the most suitable strain and expression conditions including concentration, temperature and time of IPTG induction (1.0 mmol/L, 30℃ and 2 h). Recombinant protein was purified by the means of His-tag-specific affinity choromatography after expressed. Polyclonal antibody against 2S was generated by immunizing New Zealand rabbits with purified recombinant protein 2S-His. ELISA results indicated that the two rabbits titer of anti-serum was up to 1:25 600 and 1:12 800 respectively. In conclusion, anti-serum, which could be used to detect spider dragline silk protein in vitro,was successfully obtained from 2S-pET-52b(+). This research laid basis for establishing method of expression spider dragline silk protein gene in hair.
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