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异亮氨酸拉链修饰的可溶性CD40L在巴斯德毕赤酵母中的表达 被引量：1

Expression of Isoleucine Zipper Modified Soluble CD40L in Pichia Pastoris
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摘要 为了获得异亮氨酸拉链修饰的可溶性CD 40L(IZ-sCD 40L)在巴斯德毕赤酵母中的分泌表达,首先用PCR和重叠PCR的方法得到了IZ-sCD 40L基因片段,然后构建了巴斯德毕赤酵母表达质粒pP ICZαA-IZ-sCD 40L,核酸测序分析表明IZ-sCD 40L的基因片段正确克隆到pP ICZαA质粒中,将其线性化后,电转化到巴斯德毕赤酵母G S115中,PCR筛选阳性重组菌株,经表型鉴定后,用甲醇进行诱导表达,经SDS-PAGE和W estern b lot印迹杂交结果证实了表达产物为重组IZ-sCD 40L的融合蛋白。 To obtain the expression of Isoleucine Zipper modified soluble CD40L （IZ-sCD40L） in Pichia pastoris, firstly, DNA fragment of IZ-sCD40L was obtained by PCR and over-lap PCR . Then the expression vector pPICZaA-IZ-sCD40L was constructed. Nucleotide sequencing analysis indicated that the DNA fragment of IZ-sCD40L was correctly inserted into the pPICZaA vector. Linearized pPICZaA-IZ-sCD40L was introduced into Pichia pastoris GS115. Positive clone was selected by PCR and its phenotype was determined. The positive clone was introduced with methanol. The results of SDS-PAGE and Western blot showed that product was recombinant Isoleucine Zipper modified soluble CD40L fusion protein.
作者 杜小波 田聆 王永生 刁鹏 王国庆 魏于全
机构地区 四川大学华西医院人类疾病生物治疗国家重点实验室肿瘤中心
出处 《生物医学工程学杂志》 EI CAS CSCD 北大核心 2006年第4期844-847,共4页 Journal of Biomedical Engineering
基金 国家自然科学基金资助项目(30270524)
关键词 可溶性CD40L 异亮氨酸拉链 巴斯德毕赤酵母 表达 Soluble CD40L Isoleucine Zipper Pichia pastoris Expression
分类号 R346 [医药卫生—基础医学]
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参考文献12

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同被引文献26

  • 1张义浜,施立楠,唱韶红,巩新,熊凌霜,吴军.人sTNFR II-IgG Fc融合蛋白在毕赤酵母菌中的表达及其产物分析[J].细胞与分子免疫学杂志,2007,23(6):515-519. 被引量：7
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生物医学工程学杂志
2006年 第4期

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