Metabolites-BOC Sciences

Gene Overexpression Service

Gene Overexpression Service

A highly productive method for secondary metabolite fermentation has been committed to strain improvement. As an expert in advanced technology of overexpression, BOC Sciences provides gene overexpression services to meet our customers' needs, including overproducing target metabolites in various microbial systems and accelerating the transformation from primitive strains to industrialized strains. In particular, we have successfully utilized the yeast systems to overproduce target compounds in many of our projects.

Introduction

Microbes are known to produce a wide range of secondary metabolites, such as antibiotics, mycotoxins, enzymes and other valuable compounds. Thus, microbial fermentation has been widely used in pharmaceutical, food, chemical, and environmental protection industries. The discovery of a high-producing strain is the prerequisite for efficient metabolites production at an industrial level. Certain molecular approaches for strain improvement can directly affect the expression of early metabolites biosynthetic genes. For instance, overexpression of specific genes encoding certain enzymes which are directly involved in the biosynthesis of target metabolites could regulate the production level of these metabolites. Increased production of the target metabolite can be achieved either by overexpressing positive regulatory genes or disrupting negative regulatory genes.

Application of Gene Overexpression

Overexpression as a genetic tool

A simple increase in the copy number of entire chromosomes or portions of chromosomes, independent of discrete mutations within genes, could cause mutant phenotypes.

Overexpression as a screening tool

For example, the availability of multicopy plasmid vectors for yeast Saccharomyces cerevisiae allows the selective amplification of individual genome segments. Increased dosage of particular genes results in overproduction of specific gene products. It thereby confers resistance to certain metabolic inhibitors, which can be used to isolate recombinant clones and identify phenotypes in wild-type cells.

Our Advantages

  • Over 20 years of experience in traditional strain breeding and fermentation production.
  • Advanced technologies in strains development.
  • Comprehensive strain expression systems cover bacterial and eukaryotic systems.
  • Developed fermentation upstream processing.
  • Cost-effective and high-quality products.
  • Advanced strain culture technology.
  • Optimized medium selection, biomass control and product induction.

Our Methods

Our rich overexpression experience in the yeast system has expanded different microbial overexpression systems, including bacterial and eukaryotic organisms. Our comprehensive strain development capacities cover the overall process of strain breeding, separation and purification.

Methods of regulating gene expression: gene expression regulation can occur at any step, including transcription of DNA into RNA and post-translational modification of proteins.

  • Transcriptional regulation consists of transcription factors and other proteins that work in concert to fine-tune the amount of RNA produced through various mechanisms.
  • Post-transcriptional regulation is the control of gene expression at the RNA level.

Methods of measuring gene expression

  • Fluorescence in situ hybridization (FISH): a specific nucleotide sequence of interest is bonded with a fluorescent probe of the single-strand DNA or RNA, and then detected and localized by fluorescence microscopy.
  • Reverse transcription polymerase chain reaction (RT-PCR): reverse RNA transcription into complementary DNA and amplification of specific DNA targets using polymerase chain reaction (PCR), which is mainly used to measure the amount of a specific RNA.
  • Northern blot: extraction of total RNA from cells and isolation of RNAs with a poly(A) tail, then RNA samples are separated by gel electrophoresis. The detection of these RNA samples is used to study gene expression.

Project Workflow

  • Customer advisory
  • Project discussion
  • Overexpression service
  • Selection of appropriate strains
  • Evaluation of the novel strain
  • Project delivery

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