Chicken anemia virus apoptin fusion recombinant protein as well as preparation method and application thereof

Abstract

The invention discloses a chicken anemia virus apoptin fusion recombinant protein as well as preparation method and application thereof. The chicken anemia virus apoptin fusion recombinant protein is prepared by the following steps of: sequence assembly and optimization, recombinant plasmid establishment and strain transformation, inducible expression of recombinant apoptin and expression product purification and solubility analysis. The invention also provides an application of the chicken anemia virus apoptin fusion recombinant protein in the preparation of antitumor drugs. The fusion recombinant protein msyB-Apoptin-TAT is subjected to efficient soluble expression, generates over 60% of inhibition rate on the growth of mantle cell lymphoma JEKO-1 and REC-1 cells and does not cause obvious inhibitory effect on human umbilical vein endothelial cells (HUVEC) while the expression product has specific antitumor activity.

Description

technical field [0001] The invention belongs to the technical field of bioengineering, and in particular relates to a chicken anemia virus apoptin fusion recombinant protein and a preparation method and application thereof. Background technique [0002] Apoptosis is a programmed process that plays an important role in development, differentiation and maintenance of the body. The occurrence and development of tumors are related to the excessive proliferation of cells and the reduction of apoptosis. Apoptin is a functional protein encoded by chicken anemia virus (CAV), which can induce the apoptosis of infected target cells. Subsequent studies have found that apoptin can selectively induce tumor cell apoptosis, but has no damaging effect on normal diploid cells. [0003] In addition to its broad-spectrum anti-tumor properties, apoptin has other interesting advantages: when it induces tumor cell apoptosis, it does not depend on the mediation of p53, so that tumor cells will no...

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Problems solved by technology

However, the patent uses pET-28a to induce expression, which requires at least 8 hours of induction time, and the induced product is an inclusion body, and the expression level is not high. In order to obtain active recombinant apoptin, it needs to go through tedious refolding and denaturation, etc. steps to complete the subsequent protein purification, which increases the cost of preparation, and it is easy to cause degradation of the recombinant protein during repeated operations

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