蜈蚣候选毒素Kappa-scoloptoxin-Ssm1c的表达与纯化
首发时间:2018-11-19
摘要:目的 通过原核表达与亲和纯化的方法,获取较高产量的少棘蜈蚣候选毒素,便于后续对其结构与功能进行研究。方法 利用Uniprot蛋白数据库筛选出功能未知的少棘蜈蚣候选毒素多肽,采用无缝克隆技术构建重组表达载体进行表达。采用Ni-NTA柱层析及反相高效液相色谱(RP-HPLC)分离纯化候选毒素,并采用质谱检测其分子量。结果 构建了稳定的重组表达质粒,分离纯化了Kappa-scoloptoxin-Ssm1c候选毒素,质谱检测其分子量为6072 Da。结论 成功表达并纯化了Kappa-scoloptoxin-Ssm1c候选毒素,为后续的结构与功能研究奠定了基础。
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Expression and Purification of Kappa-scoloptoxin-Ssm1c
Abstract:OBJECTIVE Prokaryotic expression and affinity purification were employed to obtain affluent candidate centipede toxin and facilitate the subsequent research about its structure and function.METHODS The Uniprot protein database was accessed to sift the unexploited centipede toxin peptide from Scolopendra Subspinipes Mutilans.The recombinant expression vector was constructed with seamless cloning tactic to express the fusion protein. The fusion protein was then purified by Ni-NTA column chromatography and reverse-phase high performance liquid chromatography (RP-HPLC). The molecular weight was identified by MALDI-TOF mass spectrometry. RESULTS The stable recombinant expression plasmid of the candidate toxin Kappa-scoloptoxin-Ssm1c was constructed. The fusion protein was expressed, isolated and purified successfully. The molecular weight was 6072 Da, demonstrated by MALDI-TOF mass spectrometry. CONCLUSION The Kappa-scoloptoxin-Ssm1c candidate toxin was expressed and purified successfully. The research lays the foundation for further study of the structure and function of Kappa-scoloptoxin-Ssm1c.
Keywords: Bioengineering Scolopendra subspinipes mutilans Prokaryotic expression Protein purification
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蜈蚣候选毒素Kappa-scoloptoxin-Ssm1c的表达与纯化
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