Prokaryotic Expression and Polyclonal Antibody Preparation of Intended Spider Dragline Silk-like Peptide Gene Monomer（S）

Abstract

Abstract: This research, through prokaryotic expression to get intended spider dragline silk protein to prepare specific antibody, would lay the foundation for detecting its expression in eukaryotic cell. Using plasmid pGEX-2T as original vector, we constructed the prokaryotic expression vector pGEX-S into the MCS（multiple clone sites）of an artificial synthesized spider dragline silk protein gene inserted. Then we induced the expression of the recombinant plasmid pGEX-S in Escherichia coli competent cells B21 with the utilization of IPTG. Recombinant protein was purified by the means of GST-tag-specific affinity to gain S-GST protein. The specific antibody was prepared by immunizing mouse CDⅠ. After immune collect serum, ELISA was employed. The expression vector pGEX-S coding sequence is right after analysis. The western blot show the recombinant protein band appears at 41 kD. ELISA results indicated that two of mouse A and F have better immunoreactivity. The study proved the successful preparation of specific polyclonal antibody through prokaryotic expression.

Key words: Intended spider dragline silk protein gene monomer（S）, Prokaryotic expression, Polyclonal antibody

Meng Fanhua, Fang Jun, Wang Haitao, Xing Yanping, Qi Yu, Zhou Huanmin. Prokaryotic Expression and Polyclonal Antibody Preparation of Intended Spider Dragline Silk-like Peptide Gene Monomer（S）[J]. Biotechnology Bulletin, 2013, 0(3): 134-138.

References

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