Rekombinasi Gen Penyandi b-xilosidase asal Geobacillus Thermoleovorans IT-08 dalam Plasmid Phis1525
Abstract: Plasmid pHIS1525 is
a shuttle vector for E. coli/ B. megaterium, which contains a signal peptide
for secretion of heterologous proteins, and also equipped with a His-tag
encoding sequence. The aim of this research was to recombine the β-xylosidase
encoding gene from G. thermoleovorans IT-08 into plasmid pHIS1525. The
β-xylosidase encoding gene (xyl) was amplified by PCR method from pTP510, a
recombinant plasmid containing xylanolytic enzyme encoding gene from G.
thermoleovorans IT-08. The xyl gene was inserted into plasmid via MCS of
pHIS1525 between SacI and SphI restriction sites and propagated in E. coli
DH10β. The recombinant plasmids were analyzed by restriction analysis and
sequencing. This research showed that the xyl gene was successfully amplified
from pTP510 and recombined into plasmid pHIS1525 in E. coli DH10β. The sequence
of the inserted gene contained a signal peptide and His-tag encoding sequences.
The recombinant plasmid was called pSMX.
Penulis: S ri Sumarsih, Ni
Nyoman Tri Puspaningsih, Ami Soewandi JS
Kode Jurnal: jpbiologidd110054
