Skip to main content

Posts

An Efficient Genome-Wide Fusion Partner Screening System for Secretion of Recombinant Proteins in Yeast.

Rarely issued to produce recombinant proteins in the yeast Saccharomyces cerevisiae, we developed a new filtration system optimal genome-wide translational fusion partner (TFP), which involves the secretion signal optimal recruitment and fusion partner.  A TFP library constructed from genomic and Multi-species Recombinant Proteins  cDNA libraries were cut using a technique based invertase signal sequence trap. The efficiency of the system was demonstrated using two rarely secreted protein, human interleukin (HIL) -2 and Hil-32. TFPs optimal for the secretion-2 and Hil Hil-32 very easily, resulting in secretion of this protein to hundreds mg / L. In addition, many expressed yeast secretion signal and fusion partners are identified, leading to the efficient secretion of recombinant proteins. TFPs been found to be useful for a hypersecretion of other recombinant proteins in yields of up to several g / L. This screening technique could provide a new method for the production of various typ
Post a Comment
Read more
Recent posts

Characterization of a panARS-based episomal vector in the methylotrophic yeast Pichia pastoris for recombinant protein production and synthetic biology applications.

BACKGROUND production of recombinant proteins in the methylotrophic yeast Pichia pastoris largely depends on integrative vector. Although the stability of the integrated expression cassette was well rewarded for most applications, the availability of reliable episomal vector to host this would represent a useful tool to accelerate cloning and high-throughput screening, ameliorating also relatively high clonal variability reported in the transformants of integrative vector caused by the integration of off- targets in P. pastoris genome. Hamster Recombinant Proteins More recently, heterologous and endogenous autonomously replicating sequences (ARS) have been identified in P. pastoris by genome mining, opening up the possibility of expanding the toolbox is available for efficient episomal plasmid insert. The purpose of this technical report is to validate the 452-bp sequence ( "panARS") in the context of P. pastoris expression vector, and to compare their performance to the clas
Post a Comment
Read more

Deciphering how LIP2 and POX2 promoters can optimally regulate recombinant protein production in the yeast Yarrowia lipolytica.

BACKGROUND In recent years, species of non-conventional yeasts Model Yarrowia lipolytica has received much attention because it is a useful plant cells to produce recombinant proteins. In this species, the expression vector involving LIP2 and POX2 promoter has been developed and successfully used for the production of proteins on the same results with or even higher than other cell factories, such as Pichia pastoris. General Recombinant Proteins  However, the production process involving the promoter can be difficult to manage, especially if done on a large scale in fed-batch bioreactor, because they require a hydrophobic inducers, such as oleic acid or methyl oleate. Thus, the challenge has been to reduce the burden on the hydrophobic substrate while promoting the production of recombinant proteins. One possible solution is to replace most of the inducer with co-substrates that can serve as an alternative energy source.  However, applying such an approach would require a detailed know
Post a Comment
Read more

Implications of evolutionary engineering for growth and recombinant protein production in methanol-based growth media in the yeast Pichia pastoris.

BACKGROUND Pichia pastoris is a eukaryotic expression host is widely used for the production of recombinant proteins. Adaptive evolution laboratory (ALE) has been applied in various studies in order to improve strains for the purpose of biotechnology.  In this context, the long-term Equine Recombinant Proteins  impact of the carbon source adaptation in P. pastoris has not been addressed so far. So, we did a pilot experiment to analyze the application and potential benefits of ALE to the increased growth and production of recombinant proteins in P. pastoris. RESULTS Adaptation to growth on methanol made in culture media replication in rich and minimal growth for 250 generations. the growth rate increased in the medium growth observed in the population and the level of a single clone. growing populations show different levels of profit growth and trade-offs in a state of non-evolutionary growth. genome resequencing reveals the potential genetic targets associated with improved growth per
Post a Comment
Read more

Bioreactor-Scale Strategies for the Production of Recombinant Protein in the Yeast Yarrowia lipolytica.

Recombinant protein production is a multibillion-dollar market. Therefore, an important area of ​​research both in academia and industry. The use of yeast as a cell factory presents some advantages such as ease of genetic manipulation, growth at high cell density, and the possibility of post-translational modifications. Yarrowia lipolytica is regarded as one of the most attractive host for its ability to metabolize the cru de substrates, Chicken Recombinant Proteins  to express the gene at high levels, and to secrete proteins in large quantities. In recent years, several reviews have been dedicated to genetic tools developed for this purpose. Although the construction of efficient cell factories for the synthesis of recombinant proteins is important, the development of efficient processes for the production of recombinant proteins in a bioreactor is an equally important aspect. Indeed, the sports car can not drive fast on a gravel road.  The purpose of this study is to provide a compre
Post a Comment
Read more

The Immunological Properties of Recombinant Multi-Cystatin-Like Domain Protein From Trichinella Britovi Produced in Yeast.

Trichinellosis is a parasitic disease globally-distributed zoonosis caused by nematodes of the genus Trichinella. One of the most common species are known to affect human health Trichinella is T. britovi; However, it is relatively poorly investigated. A thorough knowledge of the protein expressed by Trichinella important when developing immunological detection methods and vaccines and study the interaction with the host.  This study using Pichia pastoris expression Caprine Recombinant Proteins  system to produce TbCLP soluble antigen that induces a strong antibody response in the host during a natural infection. Our results demonstrate the feasibility of the production of antigen TbCLP in yeast, which is able to carry out post-translational modifications such as glycosylation and formation of disulfide bonds; they also indicate that the glycosylation TbCLP antigen having immunogenic effects in mice are tested and trigger the Th1 / Th2 mixture, and is associated with reduced larval lo
Post a Comment
Read more