High-level expression of a novel liver-targeting fusion interferon with preferred Escherichia coli codon preference and its anti-hepatitis B virus activity in vivo.
High-level expression of a novel liver-targeting fusion interferon with preferred Escherichia coli codon preference and its anti-hepatitis B virus activity in vivo.
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DOI：
10.1186/s12896-015-0177-1
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发表时间：
2015-06-12
影响因子：
3.5
通讯作者：
Zhu J
中科院分区：
文献类型：
--
作者：
Lu X;Wang J;Jin X;Zhu J
在我们前期的研究中，成功构建了一种将IFN α2b与疟原虫I区肽结合的新型肝脏靶向融合干扰素（IFN-CSP）。 IFN-CSP对HepG2.2.15细胞中HBV-DNA复制有显着的抑制作用。本研究的目的集中于如何产生高水平的重组IFN-CSP及其体内抗乙型肝炎病毒(HBV)活性。根据大肠杆菌(E.coli)偏好密码子使用合成编码IFN-CSP的修饰DNA片段，并转化至大肠杆菌BL21(DE3)中进行蛋白表达。通过结合单因素实验和正交试验（L（9）（3）（4））系统地检查了诱导条件。通过蛋白质印迹分析来表征纯化蛋白的抗原性。测定体内组织分布并与天然干扰素α2b进行比较。以HBV转基因小鼠为体内模型，评价重组IFN-CSP的抗HBV作用。结果表明，大肠杆菌表达系统非常有效地生产目标蛋白。我们目前的研究首次证明，通过密码子和表达条件的优化，IFN-CSP基因可以在大肠杆菌中高水平表达。纯化的重组 IFN-CSP 在体内表现出肝脏靶向潜力和抗 HBV 活性。本研究进一步支持了IFN-CSP在肝脏靶向抗HBV药物中的应用。
In our previous study, a novel liver-targeting fusion interferon (IFN-CSP) combining IFN α2b with plasmodium region I peptide was successfully constructed. IFN-CSP has significant inhibition effects on HBV-DNA replication in HepG2.2.15 cells. The aim of the present investigation was focused on how to produce high levels of recombinant IFN-CSP and its in vivo anti-hepatitis B virus (HBV) activity. A modified DNA fragment encoding IFN-CSP was synthesized according to Escherichia coli (E. coli) preferred codon usage and transformed into E. coli BL21 (DE3) for protein expression. The induction conditions were systematically examined by combining one-factor experiments with an orthogonal test (L(9)(3)(4)). The antigenicity of the purified protein was characterized by western blot analysis. The in vivo tissue distribution were assayed and compared with native IFN α2b. HBV-transgenic mice were used as in vivo model to evaluate the anti-HBV effect of the recombinant IFN-CSP. The results showed that the E. coli expression system was very efficient to produce target protein. Our current research demonstrates for the first time that IFN-CSP gene can be expressed at high levels in E. coli through codon and expression conditions optimization. The purified recombinant IFN-CSP showed liver-targeting potentiality and anti-HBV activity in vivo. The present study further supported the application of IFN-CSP in liver-targeting anti-HBV medicines.
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影响因子：
--
作者：
Lu X;Wang J;Jin X;Huang Y;Zeng W;Zhu J
通讯作者：
Zhu J
影响因子：
1.6
作者：
Neves, FO;Ho, PL;Nascimento, ALTO
通讯作者：
Nascimento, ALTO
影响因子：
3
作者：
Naz, Farha;Asad, Mohd;Hassan, Md Imtaiyaz
通讯作者：
Hassan, Md Imtaiyaz
影响因子：
3.9
作者：
Sun, JC;Chen, M;Luo, JH
通讯作者：
Luo, JH
影响因子：
3
作者：
Kim, Chang-Kyu;Choi, Jun-Ha;Oh, Jae-Wook
通讯作者：
Oh, Jae-Wook