title: A visual method for direct selection of high-producing Pichia pastoris clones.
abstract: BACKGROUND
  The methylotrophic yeast, Pichia pastoris, offers the possibility to generate a high amount of recombinant proteins in a fast and easy way to use expression system. Being a single-celled microorganism, P. pastoris is easy to manipulate and grows rapidly on inexpensive media at high cell densities. A simple and direct method for the selection of high-producing clones can dramatically enhance the whole production process along with significant decrease in production costs.
  
  
  RESULTS
  A visual method for rapid selection of high-producing clones based on mannanase reporter system was developed. The study explained that it was possible to use mannanase activity as a measure of the expression level of the protein of interest. High-producing target protein clones were directly selected based on the size of hydrolysis holes in the selected plate. As an example, the target gene (9elp-hal18) was expressed and purified in Pichia pastoris using this technology.
  
  
  CONCLUSIONS
  A novel methodology is proposed for obtaining the high-producing clones of proteins of interest, based on the mannanase reporter system. This system may be adapted to other microorganisms, such as Saccharomyces cerevisiae for the selection of clones.
authors.affiliation: Hubei Key Laboratory of Industrial Biotechnology, College of Life Science, Hubei University, Wuhan, 430062, PR China.
authors.forename: Fan
authors.initials: F
authors.lastname: Hu
authors.forename: Xin
authors.initials: X
authors.lastname: Li
authors.forename: Jie
authors.initials: J
authors.lastname: Lü
authors.forename: Pei Hong
authors.initials: PH
authors.lastname: Mao
authors.forename: Xiang
authors.initials: X
authors.lastname: Jin
authors.forename: Ben
authors.initials: B
authors.lastname: Rao
authors.forename: Peng
authors.initials: P
authors.lastname: Zheng
authors.forename: Yu Lin
authors.initials: YL
authors.lastname: Zhou
authors.forename: Sheng Yi
authors.initials: SY
authors.lastname: Liu
authors.forename: Tao
authors.initials: T
authors.lastname: Ke
authors.forename: Xiang Dong
authors.initials: XD
authors.lastname: Ma
authors.forename: Li Xin
authors.initials: LX
authors.lastname: Ma
mesh_terms: D015870:Gene Expression; D017930:Genes, Reporter; D008828:Microbiological Techniques / Q000379:methods*; D010843:Pichia / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism*; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D014170:Transformation, Genetic
chemical_list: D011994:Recombinant Proteins