Cloning of the gene encoding Endo-β-1,3-1,4 Glucanase from Bacillus subtilis subsp. spizizenii W23 on Plasmid pMMB67EH into Escherichia coli DH5α and Escherichia coli Origami

https://doi.org/10.22146/jsv.8114

Lisa Gunawan(1*)

(1) Fakultas Teknobiologi, Universitas Surabaya, Surabaya
(*) Corresponding Author

Abstract


One of the genes encoding cellulolytic enzymes, named BSUW23_10175, from Bacillus subtilis subsp. spizizenii W23 was cloned in the plasmid pMMB67EH into Escherichia coli DH5α and Escherichia coli Origami. Target gene was amplified using polymerase chain reaction (PCR) with specific primers (For- BSUW23_10175 dan Rev-BSUW23_10175) that were used for BSUW23_10175 gene amplification. Based on the PCR and restriction analyses of the transformants plasmid, it was known that clones number 8 and 9 of the Escherichia coli Origami transformants contained the recombinant plasmid pMMB_BSUW23_10175.
Cellulolytic activity assay showed that those transformants had cellulolytic activity by indution with IPTG (isopropyl-β-D-thio-galactosidase).


Keywords


Bacillus subtilis subsp. spizizenii W23, cloning, endo-β-1,3-1,4 glucanase, Escherichia coli DH5α, Escherichia coli Origami

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References

Daftar Pustaka

Amore, Antonella, Olimpia Pepe, Valeria

Ventorino, Leila Birolo, Chiara Giangrande

and Vincenza Faraco FEMS Microbiol Lett.

Industrial waste based compost as a

source of novel cellulolytic strains and

e n z y m e s . D i a k s e s d a r i

http://pubget.com/paper/23181595/Industrial_

Kloning Gen Pengkode Endo-β-1,3-1,4 Glukanase Bacillus subtilis subsp.

Lisa Gunawan

waste_based_ compost_as_a_source_of_

novel_cellulolytic_strains_and_enzymes.

Diakses pada 11 Januari 2013.

DIBY, Paul, Kanamparambil Augusthy SAJU,

Pulikottil John JISHA, Yammanuru Ramalinga

SARMA, Aundy KUMAR and Muthuswamy

ANANDARAJ. 2005. Mycolytic enzymes

produced by Pseudomonas fluorescens dan

Trichoderma spp. Against Phytophthora

capsici, the foot rot pathogen of black pepper

( P i p e r n i g r u m L . ) . D i a k s e s d a r i

http://www.annmicro.unimi.it/full/55/diby_55

_129-133.pdf. Diakses pada 6 Mei 2012.

Chander, Ramesh Kuhad, Rishi Gupta, and Ajay

Singh. 2011. Microbial Cellulases and Their

Industrial Applications. Enzyme Research

Volume 2011 (2011), Article ID 280696.

Fruste, J. P., W. Pansegrau, R. Frank, H. Blöcker, P.

Scholz, M. Bagdasarian and E. Lanka 1986.

Molecular Cloning of the Plasmid RP4 Primase

Region in a Multi-host range tacP Expression

Vector. Gene 48, 119 – 131.

Gram, Hans Christian. 1884. The Differential

Staining of Schizomycetes in Tissue Sections

and in Dried Preparations. Fortschitte der

Medicin. 2:185-189.

Khairil. 2009. Lignoselulosa. Diakses dari

http://khairilfuture.multiply.com/journal/item/

Diunduh pada 6 Mei 2012.

Modi, HA. 2010. Regulation of Exoglucanase and

Endoglucanase Biosynthesis in Streptomyces

Species. Diakses dari : http://journal-advancesd

e v e l o p m e n t a l - r e s e a r c h . c o m / w p -

content/uploads/2010/06/Regulation-of-

E x o g l u c a n a s e - a n d - E n d o g l u c a n a s e -

Biosynthesis-in-Streptomyces-Species-Modi-

HA.pdf. Diunduh pada 6 Mei 2012.

Sambrook, J., Fritsch, E.F. and Maniatis, T. 1989.

Molecular Cloning I: A Laboratory Manual.

Cold Spring Harbor, NY: Cold Spring Harbor

Laboratory.

Soetrisno. 2008. Sumber bahan bakar hayati

langsung dari selulosa. Diakses dari :

http://www.chem-is-try.org/artikel_kimia/

kimia_material/sumber-bahan-bakar-hayatilangsung-

dari-selulosa/. Diakses pada : 6 Mei

Pecinta lingkungan hidup siklus ITS. 2010.

Mengembangkan bioetanol dari selulosa.

Diakses dari : http://siklus.lmb.its.ac.id/? p=71.

Diakses pada : 6 Mei 2012.

Daniel R. Zeigler, Zoltán Prágai, Sabrina Rodriguez,

Bastien Chevreux, Andrea Muffler, Thomas

Albert, Renyuan Bai, Markus Wyss and John B.

Perkins. 2008. The Origins of 168, W23, and

Other Bacillus subtilis Legacy Strains. Diakses

dari : http://jb.asm.org/content/190/21/6983.

abstract?ijkey=05bef 429089ec10c2d0a49

d1d7956b3c3f106bfc& keytype2=tf_ipsecsha.

Diakses pada : 10 Januari 2013.



DOI: https://doi.org/10.22146/jsv.8114

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