High-throughput Screening for Resistance Genes Using Yeast

Due to the persistent and non-degradable of heavy metals, their pollution from industrial and agricultural activities has become a worldwide concern. The accumulation of heavy metal in soil may result in decline in crop yield and quality. Moreover, heavy metals accumulated in plants and animals can enter the food chain and subsequently harm human health. Thus, development of heavy metal hyperaccumulator plants for soil remediation is an eco-friendly and cost effective remediation measure. In view of the fact that heavy metal tolerance and accumulation in plants is associated with multiple genes that are involved in diverse functions, such as metal ion absorption and transport, sequestration, chelation, detoxification, and signal transduction, a high-throughput approach for screening of these genes is needed.

On the other hand, in agriculture, the extensive usage of pesticides in the world has also led to contamination of soli as well as drug tolerance of pests. It is an important goal in many breeding projects to create plants with strong resistance to pests and pathogens by transgene. Subsequently, screening resistance genes responsible for pest and disease infection becomes increasingly important. Besides, spraying suitable biological molecules such as dsRNAi compounds to silence genes associated with weed resistance to herbicides providing an innovative way to reduce herbicides application. Thus, it is also necessary to explore herbicides resistance genes of weeds.

In addition, understanding the complex responses of organisms at the genome level to environmental stressors, drugs, and other toxicants is of paramount importance in fields ranging from environmental health to pharmacology and drug development and to biotechnology in general. However, there is a number of limitations for the toxicity mechanism research using animal models. Thus, it is urgent to develop an alternative approach to animal testing for toxicogenomics study.

Yeast (Saccharomyces cerevisiae) is a promising experimental system for high-throughput screening for resistance genes because of its inherent features:

• A unicellular non-pathogenic microorganism with rapid and inexpensive growth.
• Amenable to genetic manipulation.
• Genome-wide analyses are easily implemented.
• A strikingly high-level of functional conservation within the human genome and other higher eukaryotes.
• The functional information available for nearly every gene.

Yeast is a powerful tool for obtaining an integrated assessment and genome-wide perspective of toxicity mechanisms through the combination of other omics. It is widely used in many fields including environmental health, agriculture, drug development, and biotechnology (Figure 1).

Figure 1. The extensive application of yeast system in environmental health, agriculture, drug development, and biotechnology (dos Santos et al., 2012).

With years of experience in molecular biology, Lifeasible offers one stop services for high-throughput resistance genes screening by yeast, including:

• Construction of yeast expression library.
• Isolation of RNA.
• Construction of cDNA library.
• High-throughput screening for resistance genes.
• Preparation of yeast working solution.
• Exploring for appropriate adverse conditions.
• Screening of positive yeast clones under adverse conditions.
• Gene identification of yeast positive clones.
• Protein-induced expression and validation.
• Phenotypic validation of yeast.
• Knock out yeast strain creation and transformation.
• Complement (overexpression) verification.

Our services are featured with high reliability, short turnaround time, and competitive price. We will provide you customized protocol to meet your demand. Our technicians are available at 24 hours, welcome to contact us for further information.