J4 ›› 2013, Vol. 26 ›› Issue (2): 41-47.doi: 10.3976/j.issn.1002-4026.2013.02.008
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GUO Kai, ZHAO Xiao-Yan, ZHOU Hong-Zi, CHEN Kai, LI Ji-Shun, CHEN Quan, YANG He-Tong
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Abstract:
We obtained fulllength nucleic acid sequence of an recombinant immunotoxin of GnRH and PE39KDEL by gene synthesis based on the E. coli codon bias. Factor Xa restriction site was inserted at the C termanol of GnRHPE39KDEL to obtion pET24bGnRHPE39KDEL recombinated plasmid which was then converted into E. coli BL21 (DE3) and BL21 (DE3) plyS. Results show that recombinant immunotoxin exhibits highlevel soluble expression after 0.2 mmol/L IPTG induction at 30 ℃ for 2 h in LB media with 0.2 % glucose till OD600 is acquired at 37 ℃ for about 0.6 h. The purity of fusion protein is above 95% and the protein yield is about 2.6 mg/g after sonication, highspeed centrifugation, HisTrap column purification and desalt. IC-50 detection indicates that the recombinant protein can significantly inhibit the growth of tumor cell strains.
Key words: GnRH, PE39KDEL, soluble expression, expression strain
CLC Number:
Q753
GUO Kai, ZHAO Xiao-Yan, ZHOU Hong-Zi, CHEN Kai, LI Ji-Shun, CHEN Quan, YANG He-Tong. Soluble expression and purification of GnRHPE39KDEL and the inhibition to the activity of a tumor cell[J].J4, 2013, 26(2): 41-47.
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URL: https://www.sdkx.net/EN/10.3976/j.issn.1002-4026.2013.02.008
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