red horizontal rule

The MBL is equipped/capable of real-time PCR, Q-PCR construction of expression vector, plasmid transformation (electroporation or chemical transformation), screening and selection of production clone, Northern, Southern and Western Analysis, and electric mobility shift assay.

The MBL provides the following services: expression vector construction and sequence confirmation, transformation into expression systems and high-throughput screening of clones, optimization of culture conditions for expression at the shake flask level, and cell bank production and characterization.

Research

  • Regulation of the P pastoris AOX
  • Primary focus on physiology and molecular biology of methylotrophic yeast, Pichia pastoris.
  • P. pastoris is utilized as a system for the production of heterologous proteins (e.g., vaccines, peptide hormones, and industrial enzymes). We are interested in the regulatory sequences of the AOX1 promoter and proteins that up and down regulates the promoter. The goal is to affect its regulation, making it weaker or stronger as needed.

Capabilities

  • PCR
  • Cloning
  • Transformation
  • Screening of transformants
  • Northern, Southern and Western Analysis
  • Electric Mobility Shift Assay (EMSA), [Gel shift Assay]
  • Foot Printing Assay

Protein Expression Services

  • Pichia vector construction and sequence confirmation
  • Transformation
  • Screening for best producing clone
  • Optimization at the shake flask level

Equipment

  • Applied Biosystems PRISM 310 Genetic Analyzer
  • Applied Biosystems 7500 realtime PCR System
  • AirClean 600 PCR Workstation
  • Shimadzu 6C-17A Gas Chromatograph
  • Sorvall Legend RT Centrifuge
  • Bachman DU530 UV/Vis Spectrophotometer
  • UVP CL-1000 Ultraviolet Crosslinker
  • Innova 4000 Incubator Shaker
  • Innova 4400 Incubator Shaker
  • Sorvall Centrifuge
  • Revco Freezer (-50C to -86C)
  • Revco Freezer (1C to -30C)
  • LabConCo Centrivap Console