| 王珏张野苏雯静李颖连建奇白雪帆黄长形.白细胞介素-21 与乙型肝炎病毒前S2S 抗原融合蛋白
在293T 细胞中的表达[J].,2012,12(23):4401-4404 |
| 白细胞介素-21 与乙型肝炎病毒前S2S 抗原融合蛋白
在293T 细胞中的表达 |
| Expression of Fusion Protein of Interleukin-21 and Hepatitis B Virus Pre-S2and S Antigen in 293T Cells |
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| DOI： |
| 中文关键词: 白细胞介素-21 乙型肝炎病毒 前S2S 抗原 融合质粒构建 转染 |
| 英文关键词: Interleukin-21 Hepatitis B virus Pre-S2 and S antigen Confluent plasmid construction Transfection |
| 基金项目:国家自然科学基金（81072434）；国家" 艾滋病和病毒性肝炎等重大
传染病防治" 科技重大专项（2008ZX10002-006, 2012ZX10002-007）；唐都医院2011 年苗子人才资助计划 |
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| 中文摘要: |
| 目的：构建白细胞介素-21（interleukin-21, IL-21）和乙型肝炎病毒前S2S 抗原（S2S）的融合表达质粒，并研究其在293T 细胞
中的表达。方法：采用PCR 方法扩增IL-21 和HBV 前S2S 基因片段，分别克隆入pcDNA3 真核表达质粒，用分子克隆方法构建
融合表达质粒，并以脂质体2000 转染293T 细胞，分别应用ELISA 法和Western Blot 法检测细胞上清及细胞中IL-21 和HBsAg
的表达水平。结果：经酶切鉴定及DNA 序列证实重组质粒内插入片段序列正确，三种重组质粒分别命名为pcDNA-IL-21、
pcDNA-S2S 和pcDNA-IL-21-S2S，并且重组质粒能在293T 细胞内表达并分泌相关蛋白。结论：成功构建IL-21 和乙型肝炎病毒
前S2S 抗原的融合表达质粒，重组质粒能在真核细胞内表达。 |
| 英文摘要: |
| Objective: To construct eukaryotic expression plasmids carried mouse interleukin-21 (IL-21) and hepatitis B virus
pre-S2 and S (HBV S2S), and to investigate the expression of the plasmids in 293T cells. Methods: IL-21 and HBV S2S were amplified
by polymerase chain reaction (PCR), and then they were cloned to pcDNA3 eukaryotic expression plasmid. Recombinant plasmid
containing both IL-21 and HBV S2S were constructed by using molecular cloning. Plasmids were transfected into 293T cells by
lipofectamine 2000. Expressions of IL-21 and HBsAg in supernatant and cells were tested by ELISA and Western Blot, respectively.
Results: The sequences of insertion elements were confirmed by enzyme digestion and DNA sequencing. Three recombinant plasmids
were named as pcDNA-IL-21, pcDNA-S2S, and pcDNA-IL-21-S2S, respectively. These plasmids could be expressed and secreted to
related proteins in 293T cells. Conclusions: Recombinant eukaryotic expression plasmids containing IL-21 and HBV S2S were
successfully constructed and could be expressed in eukaryotic cells. |
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