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Protein Expression in Yeast

Pichia pastoris expression system introduction The expression system of Pichia pastoris is a rapidly developed and widely used eukaryotic expression system. It is a methanol nutrition yeast and have two Alcohol oxidase (AOX) coding genes AOX1 and AOX2. The two sequences are similar to each other, but the AOX1 gene is strictly regulated by methanol induction and regulation. When methanol is the only carbon source, the AOX1 promoter can be induced by methanol and initiate the expression of ethanol oxidase, which can be metabolized by methanol. The plasmid containing the AOX1 promoter can be used to promote the expression of the target gene which encoding foreign protein. Characteristics of expression strain GS115: Both AOX1 and AOX2 gene could express AOX protein (Alcohol Oxidase), high methanol utilization, close to the wild type, belong to methanol rapid use type (Mut+). KM71: Most AOX1 gene was knocked out, only AOX2 coding the AOX protein, the rate of using methanol is slow, belong to methanol slow use type (Muts). different integration of the exogenous gene will cause different phenotypes of expression strains: ...

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Protein Expression in Bacillus Subtilis

Bacillus subtilis expression system introduction 1. Simple cell wall composition, the secreted protein products don’t contain endotoxin. 2. The ability to produce a large number of extracellular/target proteins, simple recovery and purification. 3. Having a high efficient secretory signal peptide and molecular chaperone system, could achieve high efficient secretory of target protein, avoid the formation of inclusion bodies. Advantages of the Bacillus subtilis expression service in Leading Biology 1. Having a variety of Bacillus subtilis expressing vectors, such as pHT01 and pHT254. 2. Having many different expressions hosts, such as intracellular expression host AS1, 1012, 168, secretory expression host WB800N, etc to fulfill different requirements of customers. 3. Could prepare high efficient competent Bacillus subtilis. Service process Classic case An influenza virus capsid protein, molecular weight 100kD, the customer provides genetic information, after genetic optimization, the vector was constructed and expressed in Bacillus subtilis 168 and 1012: &nb...

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Protein Expression in E coli

Introduction Escherichia coli (E. coli) protein expression system is one of the most widely used system to produce recombinant proteins in recent decades. After developed and improved continuously by scientists for years, E. coli expression system is being able to express a wide variety of different types of recombinant proteins in many fields, even proteins that contain complex structures. Two features that make E. coli highly useful in lab research field are its rapid reproduction ability and its generation of clones, or genetically identical bacteria. This system is applied to prepare antigens, ligands, cytokines, bacterial proteins and so on. Leading Biology’s E.coli protein expression system serves you with a simple structure, clear genetic background, short culturing period and low culture cost. Escherichia coli expression system 1. Clearer mechanism of metabolic pathways and gene expression Through whole-genome sequencing, there are total 4,405 open reading frames; Mature gene cloning and expression system; Rapid propagation...

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Virus-like particles/Phage library expression system based

Introduction The so-called virus-like particles (VLP) is the artificial nanostructures produced by the virus gene modification, this kind of nanostructures could express the target protein. The VLP has self-assembly characteristics of a true virus, but it loses the Infectious ability of the virus. The gene that encodes the virus structural protein and the gene that encodes the target protein were fused together by using molecular biology technique, and the gene then transfected to a suitable host cell. The chimeric protein produced by this technique can be automatically assembled into a virus, and the target protein is expressed on the capsid of the virus. VLP The insect cell system has been proven to be a powerful system for rapidly and easily producing VLPs, due to several convenient features, including its versatility and the rapid lead time needed for the construction of recombinant baculovirus. Besides that, insect cells system has other prominent advantages such as the high growth rate, the capacity for large-scale production, and the ability of several post-translational modifications similar to mammalian cells system. The major limit...

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Cell-free Expression System

Introduction The so-called non-cell protein expression system refers to the expression of protein by using in vitro translation system (such as ribosome display technology) to mimic the protein translation system in cell, at the same time using the well-designed liposome membrane to replace the cell membrane, proteins were inserted on it to maintain the maximum natural conformation and natural activity. In vitro protein expression is the production of recombinant proteins in solution using biomolecular translation machinery extracted from cells. Because protein synthesis occurs in cell lysates rather than within cultured cells, the method is also called cell-free protein expression. Cell-free protein production can be accomplished with several kinds and species of cell extract, and these approaches have several advantages and features that complement traditional in vivo methods. Introduction to in vitro protein expression In vitro protein expression (also known as in vitro translation, cell-free protein expression, cell-free translation, or cell-free protein synthesis) is a technique that enables ...

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