Expression and purification of porcine PID1 gene in Escherichia coli

Huan WANG; Xiaoling CHEN; Zhiqing HUANG; Bo ZHOU; Gang JIA; Guangmang LIU; Hua ZHAO

Makale detayları

Expression and purification of porcine PID1 gene in Escherichia coli

Yazarlar:

Huan WANG | Xiaoling CHEN | Zhiqing HUANG | Bo ZHOU | Gang JIA | Guangmang LIU | Hua ZHAO

Yayıncı:

TUBITAK

Dergi:

Turkish Journal of Biology 2014 Vol.38 Issue 4, pp.523-527

Belge Tipi:

Makale

Yayın Tarihi:

3/2014

Belge Dili:

Türkçe

Document ID (Org No.):

2000812171

Sıklık:

İki Aylık

eISSN:

1303-6092

pISSN:

1300-0152

Anahtar kelimeler:

Özet:

 

In this study, in order to scale up the production of recombinant porcine phosphotyrosine interaction domain containing 1 (pPID1), a pET-28a (+)-pPID1 expression plasmid was constructed and transformed into Escherichia coli Rosetta (DE3). The recombinant pPID1 was then purified and identified by western blotting, and was also analyzed in vitro for its function. The recombinant protein was tagged with only a His6 tag at its C-terminus, which could be conveniently purified by affinity column. The protein could be induced for efficient expression with 0.75 mM IPTG for 8 h at 30 °C, yielding approximately 3 mg/L. In vitro biological activity assay demonstrated that the refolded purified recombinant pPID1 increased 3T3-L1 preadipocyte proliferation. This study provides a reliable technique for the recombinant expression and purification of pPID1 proteins.

 

In this study, in order to scale up the production of recombinant porcine phosphotyrosine interaction domain containing 1 (pPID1), a pET-28a (+)-pPID1 expression plasmid was constructed and transformed into Escherichia coli Rosetta (DE3). The recombinant pPID1 was then purified and identified by western blotting, and was also analyzed in vitro for its function. The recombinant protein was tagged with only a His6 tag at its C-terminus, which could be conveniently purified by affinity column. The protein could be induced for efficient expression with 0.75 mM IPTG for 8 h at 30 °C, yielding approximately 3 mg/L. In vitro biological activity assay demonstrated that the refolded purified recombinant pPID1 increased 3T3-L1 preadipocyte proliferation. This study provides a reliable technique for the recombinant expression and purification of pPID1 proteins.

Makale detayları

Expression and purification of porcine PID1 gene in Escherichia coli

Yazarlar:

Huan WANG | Xiaoling CHEN | Zhiqing HUANG | Bo ZHOU | Gang JIA | Guangmang LIU | Hua ZHAO

Yayıncı:

TUBITAK

Dergi:

Turkish Journal of Biology 2014 Vol.38 Issue 4, pp.523-527

Belge Tipi:

Makale

Yayın Tarihi:

3/2014

Belge Dili:

Türkçe

Document ID (Org No.):

2000812171

Sıklık:

İki Aylık

eISSN:

1303-6092

pISSN:

1300-0152

Porcine PID1

Escherichia coli

expression and purification

identification

3T3-L1 preadipocytes

Sayı

Makale detayları

Expression and purification of porcine PID1 gene in Escherichia coli

Yazarlar: Huan WANG | Xiaoling CHEN | Zhiqing HUANG | Bo ZHOU | Gang JIA | Guangmang LIU | Hua ZHAO

Yayıncı: TUBITAK

Dergi: Turkish Journal of Biology 2014 Vol.38 Issue 4, pp.523-527

Belge Tipi: Makale

Yayın Tarihi: 3/2014

Belge Dili: Türkçe

Document ID (Org No.): 2000812171

Sıklık: İki Aylık

eISSN: 1303-6092

pISSN: 1300-0152

Porcine PID1

Escherichia coli

expression and purification

identification

3T3-L1 preadipocytes

Sayı

Yazarlar:

Huan WANG | Xiaoling CHEN | Zhiqing HUANG | Bo ZHOU | Gang JIA | Guangmang LIU | Hua ZHAO

Yayıncı:

TUBITAK

Dergi:

Turkish Journal of Biology 2014 Vol.38 Issue 4, pp.523-527

Belge Tipi:

Makale

Yayın Tarihi:

3/2014

Belge Dili:

Türkçe

Document ID (Org No.):

2000812171

Sıklık:

İki Aylık

eISSN:

1303-6092

pISSN:

1300-0152