| Summary: | 碩士 === 嘉南藥理科技大學 === 生物科技系暨研究所 === 97 === To express the recombinant protein into periplasmic space, and furthersecret into extracellular mediumin, in E. coli, the expression vectors,pET21-YebF, pET21-YebFEZZ, pET21-6HisYeb, pET26-DsbA, pET26-DsbC,pET26-Crr were constructed and test. The results show that all fusion tag can be expressed into perimplasmic space.
Two disulfide-bond containing proteins, human epidermal growth factor(hEGF) and snake cardiotoxin A3 (CTX A3), were test in these expression systems. hEGF can be expressed into perimplasmic space in E. coli in all vectors test. The suitable vectors are those containing soluble fusion tag, pET-yebF-EZZ,pET-DsbA, and pET-DsbC. None of the bands corresponding CTX A3 fusion protein were seen in SDS-PAGE. The CTX A3 containing plasmid lost severly after the induction. It implies that the expression of CTX A3 causes the death of expression host.
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