Effect of codon optimization on expression levels of human cystatin C in Pichia pastoris.

Human cystatin C (CysC) is a cysteine proteinase inhibitor with many potential applications. To facilitate further studies of the functions and applications of CysC, we improved the heterologous expression of CysC using a basic codon optimization method. In this study, we cloned the high-GC content wild-type sequence of the CysC gene and also designed a slightly AT-biased sequence, with codons optimized for expression in the Pichia pastoris GS115 strain. Our results showed that the optimized coding sequence of human CysC increased the expression and secretion of the CysC protein by approximately 3- to 5-fold (90-96 mg CysC/L) in yeast, compared with the expression levels of the native CysC gene (17.9-18.4 mg CysC/L). We designed, constructed, and applied an optimized version of the CysC gene for the Pichia expression system. Our results demonstrate that the optimized coding sequence provides a higher yield of secreted CysC than that produced using the wild-type gene. Our data also serve as a practical example demonstrating a rational design strategy for the heterologous expression of secreted proteins.

An MPS-BNS mixed strategy based on game theory for wireless mesh networks.

To achieve a valid effect of wireless mesh networks against selfish nodes and selfish behaviors in the packets forwarding, an approach named mixed MPS-BNS strategy is proposed in this paper. The proposed strategy is based on the Maximum Payoff Strategy (MPS) and the Best Neighbor Strategy (BNS). In this strategy, every node plays a packet forwarding game with its neighbors and records the total payoff of the game. After one round of play, each player chooses the MPS or BNS strategy for certain probabilities and updates the strategy accordingly. In MPS strategy, each node chooses a strategy that will get the maximum payoff according to its neighbor's strategy. In BNS strategy, each node follows the strategy of its neighbor with the maximum total payoff and then enters the next round of play. The simulation analysis has shown that MPS-BNS strategy is able to evolve to the maximum expected level of average payoff with faster speed than the pure BNS strategy, especially in the packets forwarding beginning with a low cooperation level. It is concluded that MPS-BNS strategy is effective in fighting against selfishness in different levels and can achieve a preferable performance.