Ammonium Transport Function Yeast Assay

Service Introduction

It is specially designed for the study of yeast ammonia metabolism and the function of transporters based on the principle of yeast heterologous complementation experiment 31019b is mostly used in the identification test of ammonium transporter. This strain can grow normally in the basic medium and grow poorly in the medium lackingium ions. pDR196 is a highly efficient expression vector of yeast protein, and the screening marker is Ura3. The gene X encoding the ammonium transporter constructed into the pDR196 vector, and it is transformed into the competent cells of 31019b. The X protein will mediate the absorption ofium salt by yeast cells, so that the yeast 31019b can grow normally in the culture medium lacking ammonium salt.

Service Advantages

▶Can detect whether a protein has ammonium salt uptake transport activity, simple and efficient;

▶Our company has all the reagents, strains, and vectors required for detecting ammonium salt transport activity.

Service Process

Article Case

Identification of Regulatory Networks and Hub Genes Controlling Nitrogen Uptake in Tea Plants [Camellia sinensis (L.) O. Kuntze]

academic journal：Journal of Agricultural and Food Chemistry

Functional analysis of CsAMT1.2 in yeast. (a) Growth complementation of the ammonium-uptake-defective yeast strain 31019b by CsAMT1.2. (b) Identification of transgenic-yeast-positive clones. (c, d) Determination of the absorption rate of NH4 ⁺ in yeast cells by NMT. CK is the 31019b yeast strain with pYES2, and T1 denotes the 31019b yeast strain with pYES2:CsAMT1.2.

Applicable Fields

▶Functional identification of ammonium transport proteins: Identify the in vivo transport functions of candidate ammonium transporter genes from plants, microorganisms, and animals, and determine whether the target genes possess the core activity to mediate transmembrane ammonium ion uptake. This is a key technical approach for gene function verification.

▶Plant stress physiology research: Analyze the role of ammonium transport proteins in maintaining intracellular ammonium ion homeostasis under stress conditions such as ammonium deficiency and heavy metal stress in crops; screen for crop genes that are tolerant to low ammonium or efficient in ammonium utilization, providing targets for nutrient-efficient breeding.

▶Microbial ammonium metabolism mechanism analysis: Study the ammonium transport systems of bacteria, fungi, and other microorganisms to clarify the regulatory mechanisms of ammonium transport proteins in microbial adaptation to ammonium-deficient environments and in maintaining enzyme activity (e.g., ammonium superoxide dismutase).

▶Functional screening of ammonium transporter mutants/variants: Perform functional verification of different mutants and allelic variants of ammonium transport proteins, evaluate the impact of amino acid site mutations on transport activity, and support research on protein structure-function relationships.