【1】白喉毒素无毒突变体E52G 蛋白CRM197
CRM197 is a non-toxic mutant of diphtheria toxin having a single amino acid substitution of glutamic acid to glycine at position 52. CRM197 is a well-defined protein and functions as a carrier for polysaccharides and haptens, making them immunogenic. It is utilized as a carrier protein in a number of approved conjugate vaccines for diseases such as Streptococcus pneumoniae, Haemophilus influenzae b and Neisseria meningitidis.
CRM197 is the ideal carrier protein or immunological adjuvant for conjugate vaccines. CRM197 has been used commercially in prophylactic vaccines since 1990. Current research and development areas include immunotherapies and anti-addiction vaccines. Pfenex CRM197 is similar in both structure and function to the protein produced in C. diphtheriae, as shown in physicochemical and in vivo studies.
https://www.crm197.com/
【2】白喉毒素无毒突变体H21G蛋白的分泌性表达及纯化
范锋锋,李燕婷,金鑫,刘月萍,吴朝今,冯宜扬,乔瑞洁,许博,赵志强,谭小梅,谢贵林
兰州生物制品研究所有限责任公司,甘肃省疫苗工程技术研究中心,中国生物技术股份有限公司
以p ET16a-H21G质粒为模板,PCR扩增H21G基因,亚克隆至p ET22b载体中,构建重组原核表达质粒p ET22b-H21G,转化感受态大肠埃希菌BL21(DE3),IPTG诱导重组H21G蛋白分泌性表达。目标蛋白以可溶形式存在于宿主菌周质间隙,表达量为10%~15%;经纯化后,重组蛋白纯度达95%以上,等电点在4.55~6.20之间,相对分子质量和N-末端氨基酸序列均与白喉毒素相符合,为下一步开发以重组H21G蛋白为基础的疫苗奠定了基础。
DOI:10.13200/j.cnki.cjb.001549
【3】白喉毒素E154D、E154R突变体及生物活性评价
高川,王惠芳,张靖,宋云扬,胡亚兰
北京药物化学研究所
在量子化学计算的基础上,结合目前关于白喉毒素结构与功能的研究状况,选择把白喉毒素催化区的第154位谷氨酸分别突变为天冬氨酸和精氨酸,研究此处电荷性质的改变对生物活性的影响.通过基因定点突变方法制备这两个突变体基因,并在大肠杆菌表达系统中获得高效表达,在此基础上对它们的生物活性进行了评价.结果表明,与重组野生型白喉毒素相比,突变体E154D的整体动物毒性和细胞毒性略有增加,而E154R的毒性下降.
文献:生物化学与生物物理进展,2006, 33(6): 574~583
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【wkh, 2020-05-04 17:14:28】
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