PcDNA3/sIL-1RⅠ重组载体体外表达产物生物学活性的检测  

Bioassay of the soluble human interleukin-1 receptor Ⅰrecombinant plasmid expression in vitro

作　　者：赵川江[1] 章锦才[2] 徐琛蓉[2] 张蕴惠[3] 

机构地区：[1]广州中山大学光华口腔医学院牙周黏膜科,510055 [2]广东省口腔医院牙周科 [3]四川大学华西口腔医学院牙周科

出　　处：《实用口腔医学杂志》2006年第6期751-754,共4页Journal of Practical Stomatology

摘　　要：目的:检测构建的PcDNA3/sIL-1RⅠ真核表达载体在体外的表达产物是否具有生物学活性。方法:脂质体介导PcDNA3/sIL-1RⅠ体外转染CHO细胞,MTT法检测重组质粒转染的CHO细胞上清液能否阻断IL-1β生物学活性。结果:重组质粒转染的CHO细胞上清液中表达的sIL-1RⅠ能阻断不同浓度IL-1β抑制A375细胞生长的作用。结论:PcDNA3/sIL-1RⅠ重组质粒在体外导入哺乳动物细胞后能够表达具有相应生物学活性的目的产物。Objective：To study the expression of soluble human interleukin-1 receptor Ⅰ （sIL- Ⅰ ） after transfeetion of recombinant plasmid PeDNA3/sIL-1R Ⅰ into CHO cells and to evaluate the bioactivity of the expressed sIL-1R Ⅰ . Methods：CHO cells were transfected with recombinant plasmid PcDNA3/sIL-1R Ⅰ or empty vector by liposome. The expression of sIL-1R Ⅰ in the cells was examined by RT-PCR. Supernatant of the cultures of sIL-1R Ⅰ or empty vector transfected cells was respectively collected. A375 cells were euhured by 50% supernatant with the presence of 0. 125- 1. 000 μg/L IL-1β, The proliferation of A375 cells was evaluated by MTT assay. Results： sIL-1R Ⅰ was found in the recombinant plasmid PeDNA3/sIL-1R Ⅰ transfeeted CHO cells and not in the empty vector transfected cells. The proliferation of A375 cells was significantly inhibited by the supernatant of sIL-1R Ⅰ transfeeted cell culture with the presence of IL-1 β （ P 〈 0.05 or P 〈 0.01 ）. Conclusion ： Recombinant plasmid PcDNA3/sIL-1R Ⅰ can be expressed in mammalian cells and may antagonize IL-1 β induced cell proliferation.

关 键 词：白细胞介素1受体 重组质粒 基因表达 生物学活性 

分 类 号：Q78[生物学—分子生物学]