Construction of a recombinant plasmid containing xynb gene from Bacillus subtilis subsp. spizizeniiW23
| Main Authors: | Wahjudi, Mariana , ., Catherina, Daniel, Xavier |
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| Format: | Proceeding PeerReviewed application/pdf |
| Terbitan: |
, 2014
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| Subjects: | |
| Online Access: |
http://repository.ubaya.ac.id/24580/1/international%20seminar%20on%20biotechnology_2014.pdf http://repository.ubaya.ac.id/24580/ |
Daftar Isi:
- This study aimed to clone thexynB gene from Bacillussubtilissubsp. spizizeniiW23,encodinga xylan l,4-beta-xylosidase to pMMB67EH plasmid which then be used to transformed EscherichiacoliDH-5a and Origami host cells. The xynBgenewas amplifiedby polymerasechain reaction (PCR) technique using a pair of primers flanking the gene sequence,and chromosomal DNA of the W23 strain as a template. Analyses of the recombinant plasmid were done by restriction analyses, and PCR detection. The result showed that the xynB has been cloned on pMMB67EH vector. The recombinant plasmid contained the xynB gene which was confirmed by restriction analyses and by PCR detection using primers pair's specific for the xynB gene and for the vector. The xylanaseactivity of xynB gene in E.coliDH-5a and Origami host cells was assayedon luria-Bertani-xylan plate qualitatively with addition of isopropyl B-D-thio-galactoside (IPTG) as an inducer.Uponsprayingwith Congored, the cells bearing the pMMB-xynB recombinant plasmid showed a xylan-degrading activity by the appearance ofclear zanearound the colonies whilethe transformant bearing an empty plasmid showed no clear zone. It could be concluded that the cloning Arocess was succeeded.