<![CDATA[A Gene encoding -L-arabinofuranosidase (abfa) that had been cloned into recombinant E. coliDH5a/pTP510 was subcloned into Saccharomyces cerevisiae yeast system. The aim of subcloning of abfA gene intoS. cerevisiae is to express the thermostable -L-arabinofuranosidase (AbfA) enzyme in the host that is often termedas Generally Recognized As Safe (GRAS), so that this enzyme earn the broader application like in food and beverageindustries. The gene of abfA was subcloned by the amplification of Polymerase Chain Reaction (PCR) from plasmidpTP510 templat. A pair of primers, pFSacI-Af (forward) and pXhoI-Af (reverse) from designed this research wasused for the amplifcation. The PCR condition was performed as follows : beginning denaturation at 94oC for 5 min;PCR cycle 30 times that consisted of denaturation (94oC for 1 min), annealing (55oC for 30 s), andpolymerization/extension at 72oC for 2 min; and than final extension at 72oC for 7 min. The abfA gene, resulted wasinserted between GAL1 promoter and CYT1 terminator in the pYES2 expression vector. This ligation product wastransformed into E. coli TOP10 host. Restriction analysis of recombinant plasmid from this construction, thedesignated as plasmid pY-Af, showed the expected size, about 7,4 kb, which was the summation of sizes of parentalplasmid ( 5,9 kb) and fragment of DNA insert (1,5 kb).]]>
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