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| Analysing the Activity of Human β Defensin 3 and Plant des-pGlu1-Brazzein and Optimizing Condition for Expression of Target Protein Used Lactose as an Inducer |
| Chun-li li hui Ruan |
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Abstract The expressed protein had been purified and analyzed. at the same time, The inductive conditions used lactose as inducer for the flask-shaking of E.coli BL21-pET-hBD3-Bra had been optimized. The purified hBD3-Bra（recombinant protein of human β defensin 3 and des-pGlu-brazzein ）had weak antimicrobial activity, but shew little sweet . After digested by thrombin and purified by affinity column, the recombinant hBD3 had high antimicrobial activity and the natural des-pGlu1-Brazzein was 600 times sweeter than that of sucrose. The Influences of three factors which were lactose concentration, induction time and temperature on growth of strain and on yield of hBD3-Bra was analyzed in detail. The result indicated that higher lactose concentration inhibit the growth of strains（P<0.01）, but had little effect on expression of target protein among 0.5%-5%(P>0.05), Biomass would be improved as time passed（P<0.01），but the yield of target protein didn’t increase obviously simultaneously（P>0.05). Temperature was a important factor effect on growth of strain and expression of target protein（P<0.01）. Further analysis shew that the best temperature for growth was 30℃-32℃ and for expression was 30℃. Lactose as inducer was as good as IPTG as inducer (P>0.05).
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Received: 14 April 2008
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Corresponding Authors:
Chun-li li
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| [1] |
WANG Zhong-Liang, HU Yue, YU Jian-Feng, HEN Jue, MA Li-Chen, CHEN Chi-Chi, YAO Wen, GU Zhi-Liang. Prokaryotic Expression, Purification and Specific Antibody Preparation of Chicken (Gallus gallus) HNF4α Protein[J]. 农业生物技术学报, 2019, 27(5): 927-935. |
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