PURPOSE The objective of this study was construction of recombinant hEGF-pPIC9 which may be used for expression of recombinant hEGF in following studies. METHODS EGF cDNA was purchased from Genecopoeia Company and used for PCR amplification. Prior to ligation, the PCR product and pPIC9 vector was digested with EcoRI and XhoI and ligated in pPIC9 vector and subjected to colony PCR screening an...

PURPOSE Various cytokine regulates hematopoesis; they promote number of stages in stem cells biology such as proliferation, differentiation and endurance. Biological effects of SCF, as a hematopoietic cytokine; is triggered by binding to its ligand c-kit. Potential therapeutic applications of SCF include hematopoietic stem cell mobilization, exvivo stem/progenitor cell expansion, gene therapy, ...

lysates of the recombinant P. pastoris strains KM71 (pPIC9-MYO)-MutS and GS115 (pPIC9-MYO)-Mut+, which contained the integration of the myostatin gene to chromosomal DNA, was completed. When Taq DNA polymerase was used for amplification, we observed either no or nonspecific amplification products. The positive control sample yielded the 831-bp specific fragment. When the HOTStar Taq DNA polymer...

introduction: follicle stimulating hormone (fsh) is one of the pituitary glycoproteines that it consists of two subunits alpha and beta. the beta subunit is responsible for the biological activity of fsh. the aim of present study was isolation of the β subunit coding sequence containing its signal sequence from human genome and then cloning of the isolated sequence in ppic9 shuttle vector under...

The yeast-like fungus Aureobasidium pullulans PL5 is a microbial antagonist against postharvest pathogens of fruits. The strain is able to produce hydrolases, including glucanases, chitinases and proteases. The alkaline serine protease gene ALP5 from A. pullulans was cloned, inserted into the vector pPIC9 to construct pPIC9/ALP5, and then expressed in Pichia pastoris strain KM71. ALP5 had a mol...

lipases are versatile biocatalysts with a wide range of application in food, dairy, leather, paper, pharmaceutical and detergent industries.  in this study, mesophilic lipase gene from an indigenous bacillus pumilus f3 that already its gene had been sequenced and  identified, was cloned and expressed in methylotrophic yeast pichia pastoris and codon translation of lipase gene was evaluated in p...

The methylotrophic yeast Pichia pastoris was used to produce the recombinant Pisum sativum defensin (rPsd1), a small peptide from pea seeds that has a high level of antifungal activity. The plasmid rPsd1/pPIC9 was integrated into the yeast genome and methanol was used to induce expression and secretion of the recombinant Psd1, at 30 C in a fed-batch mode. The effects of different pH conditions ...

BACKGROUND Antimicrobial peptides (AMPs) are the first line of host immune defense against pathogens. Among AMPs from the honeybee Apis mellifera, abaecin is a major broad-spectrum antibacterial proline-enriched cationic peptide. RESULTS For heterologous expression of abaecin in Pichia pastoris, we designed an ORF with HisTag, and the codon usage was optimized. The gene was chemically synthet...

Soybean root nodules possess a developmentally regulated acid phosphatase (ACP) that exhibits the highest specificity for purine 5'-nucleoside monophosphates. The enzyme is a glycosylated dimer of 28- and 31-kDa subunits, which appear to be products of the same gene but differ in posttranslational modifications. In order to perform directed mutagenesis and more extensive biochemical characteriz...