In this study, bovine lactoferrin (bLf), an iron-binding glycoprotein considered an important nutraceutical protein because of its several properties, was expressed in Pichia pastoris KM71-H under AOX1 promoter control, using pJ902 as the recombinant plasmid. Dot blotting analysis revealed the expression of recombinant bovine lactoferrin (rbLf) in Pichia pastoris. After Bach fermentation and purification by molecular exclusion, we obtained an expression yield of 3.5 g/L of rbLf. rbLf and predominantly pepsin-digested rbLf (rbLfcin) demonstrated antibacterial activity against Escherichia coli (E. coli) BL21DE3, Staphylococcus aureus (S. aureus) FRI137, and, in a smaller percentage, Pseudomonas aeruginosa (Ps. Aeruginosa) ATCC 27833. The successful expression and characterization of functional rbLf expressed in Pichia pastoris opens a prospect for the development of natural antimicrobial agents produced recombinantly.
2 October 2003 in Theoretical and Applied Genetics
In recent years, agricultural crops have increasingly been attacked by more destructive insect pests, forcing modern farming to depend mainly on chemical insecticides. Although valuable, their widespread and intensive misuse has raised serious concerns about environmental and public health impacts. RNAi has been proposed as a safer alternative due to its high specificity, adaptability, and low ecological footprint. So far, dsRNA has proven effective in controlling various pest species, either through topical application or via genetically modified plants. Despite advances, large-scale implementation of RNAi remains challenging due to technical and biological hurdles that contribute to inconsistent performance. Key aspects such as dsRNA design, delivery techniques, and cellular uptake mechanisms still require refinement. Additionally, ensuring environmental stability, addressing biosafety concerns, and developing cost-effective production methods are essential for its practical application. In this review, we explore recent advances in the design and implementation of dsRNA, as well as the strategies that could support the successful integration of RNAi technology into pest management programs.
Maize is the most important crop around the world and it is highly sensitive to abiotic stress caused by drought, excessive salinity, and extreme temperature. In plants, trehalose has been widely studied for its role in plant adaptation to different abiotic stresses such as drought, high and low temperature, and osmotic stress. Thus, the aim of this work was to clone and characterize at molecular level the trehalose-6-phosphate synthase (TPS) and trehalose-6-phosphate phosphatase (TPP) genes from maize and to evaluate its differential expression in maize seedlings under drought stress. To carry out this, resistant and susceptible maize lines were subjected to drought stress during 72 h. Two full-length cDNAs of TPS and one of TPP were cloned and sequenced. Then, TPS and TPP amino acid sequences were aligned with their homologs from different species, showing highly conserved domains and the same catalytic sites. Relative expression of both genes was evaluated by RT-qPCR at different time points. The expression pattern showed significant induction after 0.5 h in resistant lines and after two to four hours in susceptible plants, showing their participation in drought stress response.