Phi C31 integrase polyclonal antibody and its use

Abstract

The invention belongs to spheres of biotechnology and gene analysis treatment technique, which specifically includes a polyclonal antibody of ªÁC31 integrating enzyme and its process for preparing and its application. The said ªÁC31 integrating enzyme is obtained by the following steps: first, expressing and purifyingªÁC31 protein, then directly mixing the purified ªÁC31 protein and the adjuvant, finally, injecting the immune animals such as rabbit, little mouse, large mouse and so on. The said polyclonal antibody can be used for specifically distinguishing theªÁC31 integrating enzyme protein of 293 human cell expressed by plasmidby, also can be used for detecting distribution of the ªÁC31 protein, or can be used for enclosing external activity of ªÁC31 integrating enzyme.

Description

technical field [0001] The invention belongs to the technical field of biotechnology and gene therapy, and specifically relates to a functional expression and purification method of ΦC31 integrase, a corresponding polyclonal antibody and its application. Background technique [0002] Phage ΦC31 integrase is a member of the serine recombinase family. In mammalian cells, ΦC31 integrase can site-specifically integrate foreign nucleotide sequences into specific sites in the genome. In recent years, it has become a Useful tool for transgenic site-specific recombination manipulation and gene therapy. [0003] In 1998, Smith et al. used the ammonium sulfate precipitation method to purify prokaryotically expressed ΦC31 integrase of about 68KD for the first time, which can bind to specific nucleotide sequences attP, attB, attL, and attR. Gel drift experiments proved that ΦC31 integrase can mediate irreversible recombination reaction. Mutation analysis showed that the serine mutatio...

AI Technical Summary

Problems solved by technology

The integration mechanism, cellular localization and toxicity of ΦC31 integrase in mammalian cells have not yet been elucidated

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