基因工程变链菌防龋疫苗的研制 |
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引用本文: | 赵小琳 宋昌玲. 基因工程变链菌防龋疫苗的研制[J]. 微生物学免疫学进展, 1998, 26(3): 10-13 |
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作者姓名: | 赵小琳 宋昌玲 |
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作者单位: | 卫生部长春生物制品研究所 |
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摘 要: | 本文采用我们自己构建的基因工程龋齿疫苗菌株,大罐发酵培养3批、菌体收量平均湿重为2.67g/L,表达率为44.27%。菌体经超声裂解,硫酸铵和链霉素粗提,再经吸附层检,凝胶过滤和离子交换柱层析等步骤纯化精提GTF抗原3批。提纯的GTF抗原,平均比活,提纯倍数和收率,分别为58.34u/mg、6.08倍和19.25%。此抗原经SDS-PAGE电泳,分子量为55KD,纯度达到96.7%,并对湿度具有较好的稳定性能。且此抗原制备的疫苗免疫家兔具有较好的免疫原性
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关 键 词: | 防龋疫苗 变链菌 基因工程 |
Preparation of Recombinant Vaccine Against Dental Caries of Streptococcus Mutans |
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Abstract: | In this study,the recombinant dental caries vaccine strain constituted by the authors was cultured in mobile pilot plant fermentor containing 30 liters of LB media.The yield of the microorganism (mean wet weight) was 2.67g/L,the expression level reached 44.27%.Havested bacterial cells were disrupted by ultrasonication.Glucosyltransferase (GTF) antigen was purified by crude extraction with ammonium suphate and streptomycin,followed by hydroxylapatite,Sephacryl S 200HR,DEAE Sepharose F.F. column chromatographies.The specific activity,purification fold and recovery of purifies GTF antigen were 58.34U/mg,6.08 fold and 19.25% respectively.Its molecular mass was 55KD.The purity reached 96.7% on SDS PAGE.This antigen possessed very good stability to temperature and it also showed better immunogenicity when rabbits were immunized with dental caries vaccine made from this antigen. |
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Keywords: | Vaccine against Dental caries Steptococcus mutants Genetic engineering Glucosyltransferase |
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