The yeast SUC2 gene, cloned on a multicopy plasmid pRB58, was used to study the effect of oxygen on the invertase expression of the recombinant Saccharomyces cerevisiae. Glucose repression was not the only factor affecting the invertase expression. The results obtained from the single-stage continuous cultures under microaerobic conditions showed that invertase expression was also strongly dependent on oxygen availability, and moving from anaerobic to aerobic conditions led to a five-fold increase in specific invertase activity. However, the cell yields under anaerobic conditions were quite low compared to those under aerobic conditions. These opposite effects of oxygen on cell growth and gene expression offer a strategy for maximizing invertase productivity by a two-stage continuous culture. The first stage was operated at a low level of glucose, around 100 mg/l, under aerobic conditions in order to obtain a high yield of yeast biomass, and the second stage maintained anaerobic conditions with residual glucose levels of 50 mg/l to derepress and fully induce invertase expression. The two-stage continuous culture resulted in a 2.5-fold increase in invertase productivity over that of a single-stage continuous culture.
Digital convergence era has been already started and is rapidly developed. Recently, convergency technology became an essential issue in all industries, and is expected to accelerate. This means that we can experience growth and change of architecture using new technology and approaching method. The AR (Augmented Reality) technology, among these convergence technology, is a kind of virtual reality technology that user can see a scene of real world which is overlapped by virtual world with additional information. It has been used in manufacturing and management in the whole industry fields including medical, mechanical, military field and so on because it can provide higher sense of reality. Thus, in this paper, we suggest the utilization of AR technology for organically connecting the roads, facilities, green area, landscape and others in the layout planning of housing complex. Users can see real world with virtual object by overlap computer graphic on the real world. This method provides users with various information about territoriality and openness. As the result of this study it is expected that this technology will help the layout planning of housing complex to become more reasonable in the aspect of design, time and cost.
We constructed a H. pylori expression vector which consisted of both a His-tag and a GST tag as purification tools for recombinant protein and a chloramphenicol resistant cat gene as a reporter. The backbone of the vector pBK contained an ColEI origin of replication and a kanamycin resistant gene. A set of oligos for the His-tag and the PCR product of gst (glutathione S-transferase) gene were inserted sequentially in frame in the multi-cloning site of pBK. The orf of cat was inserted downstream of the gst to generate pBKHGC. The 3' part of H. pylori clpB and flaA were cloned into the vector which was introduced into H. pylori. Recombinant proteins were purified by GSH affinity column, digested with thrombin and were analyzed by western blotting. The final recombinant proteins were successfully purified.
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