Cloning and Over-expression of xynB Gene of Bacillus subtilis subsp. spizizenii W23 into Escherichia coli Origami Host Cells

Wahjudi, Mariana and ., Catherina and Wangunhardjo, Nita Marcelia and Suryadjaja, Ernest and Daniel, Xavier (2017) Cloning and Over-expression of xynB Gene of Bacillus subtilis subsp. spizizenii W23 into Escherichia coli Origami Host Cells. KnE Life Sciences, 2017. ISSN 2413-0877

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Abstract

The xynB gene of Bacillus subtilis subsp. spizizenii W23 is predicted to encode a xylan 1,4-beta-xylosidase. Application of XynB enzymes in industries is wide. Production of this enzyme in its host cells is naturally restricted by repression process. It will give certain beneficial to over-expressed the enzymes in other host-cells under inducing promoter. This study aimed to clone the xynB gene from Bacillus subtilis subsp. spizizenii W23, to pMMB67EH plasmid, and to over-express the xynB gene in Escherichia coli Origami as host cells. The xynB gene was successfully amplified by polymerase chain reaction (PCR) technique using a pair of primers flanking the gene sequence and chromosomal DNA of the W23 strain as a template. The xynB gene inserted in recombinant plasmid was confirmed by PCR detection using primers pair’s specific for xynB gene and for the vector, then continued by restriction analyses. The result showed that transformants clone 9 and 10 bear the recombinant pMMB-xynB plasmid. The xylanase activity of xynB gene in Escherichia coli Origami clone 10 was detected by sodium-dodecyl-sulfate polyacrylamide gel analyses and with addition of isopropyl-

Item Type: Article
Uncontrolled Keywords: Bacillus subtilis subsp. spizizenii str. W23; cloning; Escherichia coli Origami; over-expression, xylan-beta 1,4-xylosidase (xynB).
Subjects: Q Science > QH Natural history > QH301 Biology
Divisions: Faculty of Technobiology > Department of Biology
Depositing User: Eko Setiawan 194014
Date Deposited: 21 Nov 2018 04:23
Last Modified: 21 Nov 2018 04:23
URI: http://repository.ubaya.ac.id/id/eprint/33909

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