A method for displaying foreign protein macromolecules on the surface of t4 bacteriophage using the intracellular synchronous expression method

Abstract

The invention discloses a method for showing a foreign protein macromolecule on the surface of a T4 bacteriophage by using an intracellular synchronous expression method. The method for showing the foreign protein macromolecule on the surface of the T4 bacteriophage comprises the following steps that an expression plasmid of recombinant protein fused by Hoc (Highly-antigenic Outer Capsid) protein or Soc (Small Outer Capsid) protein with a natural T4 bacteriophage control area and foreign protein is constructed; the expression plasmid of the foreign recombinant protein is transformed into an escherichia coli cell; the escherichia coli cell containing the expression plasmid of the foreign recombinant protein is infected by an Hoc protein and Soc protein deleted T4 bacteriophage virus strain; T4 bacteriophage particles generated in the infection step and showing the foreign recombinant protein are separated; and the foreign recombinant protein shown on capsid of the obtained T4 bacteriophage particles is detected. The method does not require overexpression of the foreign recombinant protein, avoids a technical difficulty that large-amount overexpression of the foreign recombinant protein is required in an in-vitro showing method, does not require separation or purification of the foreign recombinant protein, and avoids a time-consuming and expensive protein separation and purification process.

Description

Technical field

[0001] The present invention relates to the fields of biochemistry and molecular biology, in particular to a method for displaying foreign protein macromolecules on the surface of T4 phage using an intracellular synchronous expression method. Background technique

[0002] There are two modified proteins on the surface of the capsid of T4 phage, namely Hoc protein and Soc protein. These two proteins are non-essential proteins, that is, deletion or modification of these two proteins will not affect the replication and amplification of T4 phage. Based on the characteristics of the two protein molecules, and through molecular biological means, foreign proteins (that is, non-T4 phage proteins) can be recombined with Hoc or Soc proteins and displayed on the surface of T4 phage. The technology of displaying foreign recombinant proteins on the surface of T4 phage through molecular biological means has shown great potential application value in vaccine development. [0003...

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