UNIVERSITAS AIRLANGGA



Detail Article

BIOSAINS

ISSN 1412-1433

Vol. 13 / No. 3 / Published : 2011-09

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Original Article :

Recombinant gen of encoding b-xylosidase from geobacillus thermoleovorans it-08 in phis1525 plasmid

Author :

  1. S ri Sumarsih*1
  2. Ni Nyoman Tri Puspaningsih*2
  3. Ami Soewandi JS*3
  1. Departemen Kimia, Fakultas Sains dan Teknologi Universitas Airlangga
  2. Departemen Kimia, Fakultas Sains dan Teknologi Universitas Airlangga
  3. Departemen Kimia, Fakultas Sains dan Teknologi Universitas Airlangga

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.

Keyword :

β-xylosidase, Geobacillus thermoleovorans IT-08, pHIS1525, E. coli DH10β,


References :

Brown TATA,(2001) Gene Cloning and DNA Analysis: An Introduction, fourth edition USA. : Blackwell Science Ltd.,

Choi JH and SB Lee,(2004) Secretory and extracellular production of recombinant protein using Escherichia coli, - : Appl. Microbiol biotechnol

Hollmann R, M Malten, R Biendendieck, Y Yang, W Wang, D Jahn, and WD Deckwer,(2006) Bacillus megaterium as a host for recombinant protein production - : Eng. Life Sci

Jordan E, M Hust, A Roth, R iedendieck, T. Schirrmann, D. Jahn, and S. Dubel,(2007) Production of recombinant antibody fragments in Bacillus megaterium - : Microbial Cell factories

S ambrook J, and D.W. Russel (eds),(2001) Molecular Cloning: a Laboratory manual, 3rd edition New York : Cold Spring Harbor Laboratory Press





Archive Article

Cover Media Content

Volume : 13 / No. : 3 / Pub. : 2011-09
  1. The Role Of Tgf-b1 As Switching Isotype Regulator On Siga Secretion In Saliva
  2. Influent Of E. Coli And Leucocytes Againts Human Sperm Quality In Vitro
  3. Recombinant Gen Of Encoding B-xylosidase From Geobacillus Thermoleovorans It-08 In Phis1525 Plasmid
  4. The Potency Of Blue Light Emitting Diode (led) For Photoinactivation Of Staphylococcus Aureus Bacteria With Endogeneous Porphyrin
  5. The Effect Of Tomato (lycopersicum Pyriforme) Juice On The Prevention Of Photoaging Of The Skin As A Result From Ultraviolet-b Irradiation
  6. Regeneration Of Massive Bone Defect With Bovine Hydroxyapatite As Scaffold Of Mesenchymal Stem Cells

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