Position: Home > Articles > Expression of Rabbit IL-15 Protein in Pichia Pastoris Yeast and It's Immunological Activity
Journal of Shihezi University(Natural Science)
2008,26
(6)
46-49
兔IL-15蛋白在毕赤酵母中的表达及其免疫学活性
作 者：
孟庆玲;乔军;陈创夫;才学鹏;骆学农
单 位：
中国农业科学院兰州兽医研究所/家畜疫病病原生物学国家重点实验室;石河子大学动物科技学院
关键词：
兔IL-15基因;毕赤酵母;表达;免疫学活性
摘 要：
用特异性引物扩增出兔IL-15基因片段,用KpnI和NotI双酶切后将其定向克隆到pPICZαA中,构建出重组质粒pPICZαAIL-15。将pPICZαAIL-15用SacI内切酶线性化后,电转化感受态GS115酵母细胞,用PCR法筛选阳性重组子。用1.5%的甲醇诱导重组酵母菌,取培养物上清进行重组蛋白的检测。结果SDS-PAGE电泳可在重组酵母菌培养物上清中检测到分子量为18.6kDa大小的重组蛋白。凝胶薄层扫描分析表明,3株重组酵母菌在1.5%甲醇诱导144h后,重组IL-15表达量约占培养物上清总蛋白量的6.1%~8.6%。将重组IL-15用Ni离子亲和层析纯化后,在MTT试验中表现出明显的促进淋巴细胞的增殖反应,证实表达的重组IL-15具有天然蛋白的免疫学活性。
译 名：
Expression of Rabbit IL-15 Protein in Pichia Pastoris Yeast and It's Immunological Activity
作 者：
MENG Qing-ling1,2,QIAO Jun1,CHEN Chuang-fu1,CAI Xue-peng2,LUO Xue-nong2 (1 Key Lab of Agricultural Biotechnology,Shihezi University,Shihezi,Xinjiang 832003,China;2 State Key Lab of Veterinary Etiological Biology,Lanzhou Veterinary Research Institute,Chinese Academy of Agricultural Sciences,Lanzhou,Gansu 730046,China)
关键词：
rabbit IL-15;Pichia pastoris;expression;immunological activity
摘 要：
The rabbit IL-15 gene was amplified with a pair of specific primers and cloned into pPICZαA for yeast expression.The recombinant pPICZαAIL-15 was linearized with SacI and then transformed into competence GS115 for expression under the induction of 1.5% methanol.The positive yeasts were screened by PCR.The expression of IL-15 protein was detected by SDS-PAGE.The results revealed that it had a molecular weight of 18.6kDa.The expressed recombinant IL-15 protein amounted to 6.1～8.6 percent in the total protein of supernatant by analysis of gel scanning.After purification by Ni ion affinity chromatograph column,the recombinant IL-15 protein displays better stimulation to lymphocyte in MTTassay,which denotes that recombinant IL-15 protein is of better bioactivity.
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