CAJ | 학술논문

TNF与多种疾病密切相关。为了获得大量具有生物学活性的可溶性TNF受体用以拮抗TNF的毒性作用，在原核表达系统中表达了TNFR（P55）的胞外区与TrxA的融合蛋白。将TNFR（P55）胞外区去信号肽的前三个结构域基因克隆入融合蛋白表达载体pET-32a，在大肠杆菌BL21（DE3）中高效表达了TrxA-TNFR融合蛋白。表达产物以包涵体形式存在，经过变性和复性，并经镍金属鳌和柱亲和层析纯化，得到了纯度较高的可溶性受体蛋白的初纯品。免疫学实验及L929细胞体外实验均表明：该蛋白具有TNFR（P55）特异的抗原性、与TNF结合的活性以及良好的抑制TNF的生物学活性。
TNF여다충질병밀절상관。위료획득대량구유생물학활성적가용성TNF수체용이길항TNF적독성작용，재원핵표체계통중표체료TNFR（P55）적포외구여TrxA적융합단백。장TNFR（P55）포외구거신호태적전삼개결구역기인극륭입융합단백표체재체pET-32a，재대장간균BL21（DE3）중고효표체료TrxA-TNFR융합단백。표체산물이포함체형식존재，경과변성화복성，병경얼금속오화주친화층석순화，득도료순도교고적가용성수체단백적초순품。면역학실험급L929세포체외실험균표명：해단백구유TNFR（P55）특이적항원성、여TNF결합적활성이급량호적억제TNF적생물학활성。
TNF （tumor necrosis factor）is one of the most importantcytokines.In addition to its outstanding anti-tumor function and other very important functions，it is a key factor in the pathophysiology of many diseases.Soluble TNF receptor can inhibit TNF’s activity by binding to TNF but fails to transduct the signals into cells.We successfully amplified the first three domains in the extra-cellular region of TNF receptor P55 by PCR and cloned into prokaryotic expression vector pET-32a.The expressed protein was fused with TrxA and produced in the form of inclusion body.After denaturation and renaturation，we got biologically active receptor protein，which has good antigenicity and binding ability to TNF.It can also inhibit the cytotoxic activity of TNF on L929 cell culture.