Reconstructed nucleic acid construction and method for quickly recycling amino acid composition by using reconstructed nucleic acid construction

Abstract

Provided is a nucleic acid construction which is adjustable and can effectively breaks bacterial strains to release the recombinant protein in a bacterial strain cell out of the cell, thereby realizing the aim of convenient and fast recycling of the amino acid composition. The nucleic acid construction includes a nucleic acid for controlling an expressed gene and a gene product being expressed through induction; wherein the nucleic acid for controlling an expressed gene includes a heat-sensitive repressor protein) CI857 gene, a gamma PRPL promoter, a heat shock protein groEL promoter, and a lacO operator site. The gene product being expressed through induction includes a phage phiX174 lysis gene E and a phage T4 lysosome gene or a T7 lysosome gene. Taking the productions of Agrobacterium radiobacter NRRL B 1129 lamidohydrolase (AHL) and hydantoinase (HDT) as embodiments, the produced bacterial strain is provided with the nucleic acid construction, the produced bacterial strain stops growing immediately once the temperature induction is taken the effect, and the reconstructed AHL and HDT produced by induction can be effectively released out of the cell.

Description

technical field [0001] The present invention relates to a recombinant nucleic acid construct and a method for rapidly recovering amino acid composition using the recombinant nucleic acid construct, in particular to a rapid recovery of Escherichia coli from the culture medium in the industrial process for producing recombinant proteins in Escherichia coli The method of releasing the protein. Background technique [0002] The ultimate goal of the Human Genome Project is to draw out the basic blueprint of life, so that human beings can use it to understand physiological functions and spy on the mysteries of life; on the other hand, scientists can also use this map to decipher human genes The mystery, and the use of biological key technologies to solve the problem of human genetic diseases, and the development of related biotechnology to promote the future well-being of human beings. Due to the smooth promotion of this greatest human project in the 21st century, it has also pro...

AI Technical Summary

Problems solved by technology

[0007] On the other hand, Miksch et al. use the function of the kil gene to release the protein produced in the intercellular membrane (Appl.Mirobiol.Biotechnol., 1997, 47:530-536) to produce glucanase β- glucanase as an example, the results show that this technology does not cause cell rupture, but can release the glucanase produced in the intercellular membrane in the extracellular fermentation broth; however, the intercellular membrane is located in the outer and inner membranes of E. coli Because the cytoplasmic space is much larger than the intermembrane space, the protein to be produced can be accumulated and produced in a large amount in the cytoplasm compared with the intermembrane space; and the kil gene product can help the release of intermembrane proteins, which means The structure of the outer membrane of the cell is changed to cause leakage. Therefore, under the condition of huge shear stress generated by industrial fermentation scale, the cells are not easy to scale up and culture, which limits the applicability of this technology.

Images