Expression of biologically active recombinant equine interferon-gamma by two different baculovirus gene expression systems using insect cells and silkworm larvae.
Expression of biologically active recombinant equine interferon-gamma by two different baculovirus gene expression systems using insect cells and silkworm larvae.
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利用昆虫细胞和蚕幼虫通过两种不同的杆状病毒基因表达系统表达具有生物活性的重组马干扰素-γ。
作者：
Dong;K. Murakami;N. Liu;Y. Inoshima;T. Yokoyama;T. Kokuho;S. Inumaru;T. Matsumura;T. Kondo;Katsushige Nakano;H. Sentsui
将含分泌信号肽编码区的马干扰素γ (eifn - γ)全长cDNA克隆到杆状病毒转移载体pAcYM1中。将该载体与加利福尼亚自签名核多角体病毒DNA或杂交核多角体病毒DNA共转染到狐尾蛾细胞中。重组病毒，命名为aceifn - γ和hyeifn - γ，然后恢复。重组eifn - γ (reifn - γ)积累在感染了aceifn - γ或hyeifn - γ的三头虫衍生细胞株BTI TN 5B1-4的培养液和感染hyeifn - γ的家蚕幼虫的血淋巴中。这些reifn - γ形式显示为14、16、18和20kDa蛋白，并通过SDS-PAGE和tunicamycin处理证实了糖基化。通过细胞病变效应还原实验和马胎儿肾-78细胞系MHC II类抗原诱导，两种reifn - γ蛋白对水泡性口炎病毒均显示出高水平的生物活性。
The full-length equine interferon-gamma (eIFN-gamma) cDNA, including the secretion signal peptide coding region, was recloned into baculovirus transfer vector pAcYM1. This vector was co-transfected with Autographa californica nuclear polyhedrosis virus DNA or hybrid nuclear polyhedrosis virus DNA into Spodoptera frugiperda cells. The recombinant viruses, named AcEIFN-gamma and HyEIFN-gamma, were then recovered. Recombinant eIFN-gamma (reIFN-gamma) was accumulated in the culture fluid of the AcEIFN-gamma or HyEIFN-gamma infected Tricoplusia ni -derived cell line, BTI TN 5B1-4, and hemolymph of HyEIFN-gamma infected silkworm larvae. These reIFN-gamma forms were shown to be 14, 16, 18 and 20kDa proteins, and glycosylated as confirmed by SDS-PAGE and tunicamycin treatment. Both reIFN-gamma proteins, showed high-level biological activities to vesicular stomatitis virus by cytopathic effect reduction assay, and MHC class II antigen induction on the equine fetal kidney-78 cell line.
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影响因子：
4.4
作者：
J. Pace;S. Russell;P. A. Leblanc;D. Murasko
通讯作者：
J. Pace;S. Russell;P. A. Leblanc;D. Murasko
影响因子：
--
作者：
Ganes C. Sen;R. Ransohoff
通讯作者：
Ganes C. Sen;R. Ransohoff
通过单克隆抗体免疫沉淀揭示了天然人干扰素-γ的三种分子量形式。
DOI：
--
发表时间：
1984
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Kelker,HC;Le,J;Rubin,BY;Yip,YK;Nagler,C;Vilcek,J
通讯作者：
Vilcek,J
糖苷酶处理对人干扰素γ理化性质和生物活性的影响。
DOI：
--
发表时间：
1983
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Kelker,HC;Yip,YK;Anderson,P;Vilcek,J
通讯作者：
Vilcek,J