High level expression and purification of 1 C-terminal fragment of merozoite surface protein of Plasmodium falciparum in Pichia pastoris / 中国人兽共患病学报

To obtain an ideal recombinant C-terminal fragment of the merozoite surface protein of Plasmodium falciparum in the Pichia pastoris expression system, the major surface protein-119 (MSP-119) gene sequence bearing the 6-his gene was inserted into expression vector pPIC9k and the target gene was transformed to the susceptible yeast cells GS115 by using electroporation. The multiple inserts were screened and the successfully expressed MSP-119 protein with the relative molecular weight of 12kDa in the supernatants of cell cultures could be detected by SDS-PAGE. Meanwhile, Western blot analysis also demonstrated that this protein reacted with mouse anti-MSP-119 monoclonal antibody, and the expression level of MSP-119 was more than 1.0 g/L. It is concluded that this recombinant protein expressed in the Pichia pastoris expression system resembles the native proteins existed.

Construction of Human Endostatin in Yeast Eukaryotic Expression Vector / 中国普外基础与临床杂志

Objective To construct yeast eukaryotic expression vector carrying human endostatin (ES) cDNA. Methods The functional fragment of endostatin gene in human hepatic tissue was amplified by using RT-PCR technology, and cloned into yeast pPIC9 expression vector. The positive clone was sequenced by using automatized sequencer. Results The endostatin cDNA was successfully cloned. The positive ES clone gene in pPIC9 expression vector was sieved, and its coding sequence was identified to be as same as the previously reported sequence. Conclusion The successful construction of ES gene in pPIC9 expression vector using molecular biological method maybe helpful for the high expression of ES protein, which may lay the foundation for the treatment of malignant tumor through anti-angiogenesis appoach.