Production of Recombinant Protein J-Tat p-GEX Derived frm Different Re-Transformed Construct Origins  

Oleh:

Margawati, Endang Tri ; Indriawati ; Ridwan, Muhammad

Jenis: Article from Journal - ilmiah nasional
Dalam koleksi: Jurnal Mikrobiologi Indonesia vol. 11 no. 2 (Sep. 2006), page 91-94.
Topik: PROTEIN; protein

Ketersediaan

Perpustakaan Pusat (Semanggi) Nomor Panggil: JJ139 Non-tandon: 1 (dapat dipinjam: 0) Tandon: tidak ada

Lihat Detail Induk

Isi artikel

The study described here was aimed to optimize the purified recombinant protein of jembrana tat (J-Tat) in the p-GEX system. Two colonies derived from the J-Tat re-transformed construct (supernatant and pellet) were used as sources of new recombinant biral protein (J-Tat). Re transformation of J-Tat into a new cell bacterial escherchia coli was undertaken due to the old construct in old cell bacterial (E. Coli) did not express the J-Tat protein. A heat shock method was perfomed to transform the old construct into a new bacterial E. coli. In prior to transformation, the E. coli was treated in transformation storage solution to make a competent cell. Colonies of both sources were then used for trial in production of J-Tat protein in p-GEX system. Purification of solubilized protein was conducted either with resin GSTrap using a peristalic pump or altenatively using glutathione sepharose 4B in a batch capture method. Purified protein derived from supernatan performed a cleaner and a sharper band than rpotein derived from pelelt agter protein characterization by western blot. By batch capture purification with glutathione sepharose 4B, protein derived from supernatan was slightly higher concentration compared to that in pellets (i.e. 1,6 mg ml (-1) vs 1,3 mg ml (-1), respectively). For purification with resin GS trap using a peristaltic pump, both proetins of supernatant origin and of pellet origin did not show any band after western blotting. This study suggests that J-Tat recombinant protein derived from supernatan and pellets using batch capture method showed a higher yield of purified J-Tat protein.

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