Summary: | 碩士 === 東吳大學 === 微生物學系 === 92 === Alicyclobacillus acidocaldarius is a heterotrophic, saprophytic bacteria which grows in thermoacidophilic environment. Its protein can work in thermoacidophilic condition, too. The xylose isomerase gene of Alicyclobacillus acidocaldarius had been cloned and expressed in E. coli, it has great potential in food industry. Xyl operon is composed by genes of xylose isomerase, xylulosekinase, xylose peamease and regulator. In this study we clone the DNA fragment from Alicyclobacillus acidocaldarius which encode the xyl operon, and subclone it to E. coli. Then after sequencing the DNA fragment we analyzed the data in NCBI protein database in order to realize the structure of xyl operon. We got a clone, pCY10, which carries 11 kb DNA fragment of Alicyclobacillus acidocaldarius. By restriction enzyme mapping, DNA sequencing and NCBI analysis, we understood that in Alicyclobacillus acidocaldarius, the genetic organization of the xyl operon is, 5''-orf771(771 bp, predicted phosphohydrolase gene)-xylT(1410 bp, xylose permease gene)-xylA(1323 bp, xylose isomerase gene)-xylB(1536 bp, xylulose kinase gene).It is different from other bacteria. By RT-PCR test we prove that the xylAB share a common promoter, and the other genes are furn on by their own promoters.
|