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Publikasi Staf Biotek

Soluble expression and purifiation of hepatitis B core antigen (HBcAg) subgenotype B3 in Escherichia coli using thioredoxin fusion tag



Objective: To express HBcAg protein (hepatitis B virus subgenotype B3) in Escherichia coli in soluble form.
Methods: HBcAg sequence of hepatitis B virus subgenotype B3 was cloned into plasmid pET32a and introduced to E. coli BL21 (DE3). The E. coli was grown in Luria-Bertani (LB) medium supplemented with ampicillin with agitation. Protein expression was induced by adding isopropyl-β-D-thiogalactopyranoside (IPTG) at concentrations of 0.1 mmol/L, 0.3 mmol/L, and 0.5 mmol/L at room temperature (28 °C). The bacteria were dissolved in lysis buffer and lysed by freeze-thawing method then sonication. The fusion protein [thioredoxin A-(His)6tag-HBcAg] was purified using immobilized metal affinity chromatography. The protein expression was analyzed by SDS-PAGE, dot blot, and western blot.
Results: This research showed that DNA sequence of HBcAg could be propagated in pET32a and soluble protein was successfully expressed in E. coli. Induction with 0.3 mmol/L IPTG and 4-hour incubation was the best condition to express the HBcAg protein. SDS-PAGE and dot blot analysis showed that HBcAg protein could be expressed in E. coli. Western blot analysis showed that molecular weight of HBcAg fusion protein was about 38.5 kDa.
Conclusions: This study confirmed that HBcAg protein could be expressed in soluble form in E. coli.

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Judul Seri
Asian Pacific Journal of Tropical Disease
No. Panggil
-
Penerbit : .,
Deskripsi Fisik
Vol. 7 (8): 496-501.
Bahasa
English
ISBN/ISSN
-
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NONE
Tipe Isi
text
Tipe Media
other
Tipe Pembawa
online resource
Edisi
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Subyek
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