Image of Expression of No Affinity Tagged Recombinant Human Interferon Alpha-2a in Methilotropic Yeast Pichia pastoris

Publikasi Staf Biotek

Expression of No Affinity Tagged Recombinant Human Interferon Alpha-2a in Methilotropic Yeast Pichia pastoris



Recombinant human interferon alpha-2a (rhIFN╬▒-2a) has been widely used for clinical therapy as antiviral, anticancer as well as immunomodulator. In this study, the open reading frame (ORF) encoding synthetic hIFN╬▒- 2a was constructed to be in framed with N-terminal alpha factor secretion system in methylotropic yeast Pichia pastoris. This research aimed to construct, express and analyse the non-affinity tagged recombinant human interferon alpha-2a in the methilotropic yeast P. pastoris. We used pPICZ╬▒B plasmid for cloning and expression vector. The confirmed recombinant plasmid containing the correct DNA sequence of hIFN╬▒-2a was linearized by SacI restriction enzyme, then transformed into P. pastoris genome using electroporation. We screened two multi-copy recombinants in YPDS plates containing ZeocinÔäó. Buffered complex medium containing 0.5 % methanol (BMMY) was used for protein expression for 48 hours in the culture condition. The recombinant protein was purified by blue sepharose affinity chromatography. Analyses of hIFN╬▒-2a protein by SDS-PAGE and Western blot confirmed that protein band in which was observed around 19.2 kDa, was recombinant hIFN╬▒- 2a. The quantification of purified rhIFN╬▒-2a using colorimetric binichoninic assay (BCA) informed that the yield was 44 mg/L culture (OD600= 2-3).

(Ludya/Pustakawan)


Ketersediaan

Tidak ada salinan data


Informasi Detil

Judul Seri
Annales Bogorienses
No. Panggil
-
Penerbit Pusat Penelitian Bioteknologi LIPI. : Cibinong..,
Deskripsi Fisik
Vol. 19, No. 2: 57-62.
Bahasa
English
ISBN/ISSN
-
Klasifikasi
NONE
Tipe Isi
text
Tipe Media
-
Tipe Pembawa
online resource
Edisi
-
Subyek
Info Detil Spesifik
-
Pernyataan Tanggungjawab

Versi lain/terkait

Tidak tersedia versi lain


Lampiran Berkas



Informasi


DETAIL CANTUMAN


Kembali ke sebelumnyaXML DetailCite this