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无乳链球菌甘油醛-3-磷酸脱氢酶C的B细胞抗原表位预测与鉴定

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旨在表达牛源无乳链球菌(Streptococcus agalactiae)甘油醛-3-磷酸脱氢酶C(GapC)蛋白并对其免疫效果进行评估,预测并鉴定其优势B细胞抗原表位.本研究对无乳链球菌X16087株的GapC基因扩增,构建重组质粒pET-28a-WR-X16087,经诱导表达后纯化GapC蛋白,免疫小鼠并分析免疫效果.对GapC蛋白进行生物信息学分析,预测并鉴定其B细胞表位.结果表明,重组质粒经诱导表达纯化后,获得大小为44 kDa的目的蛋白,免疫小鼠后抗体效价可达1∶218700,抗体亚型以IgG1和IgG2b为主;小鼠免疫后保护率可达100%;GapC免疫后可显著减少攻击后小鼠肝、脾、肺、肾中的菌体载量;GapC蛋白可减少细菌入侵对小鼠造成病理损伤.预测并筛选了GapC 的 B 细 胞 表 位,确 定 了 3 个 具 有 良 好 抗 原 性 的 B 细 胞 线 性 表 位,分 别 为:46KYDTTQGRFDGTVEVKDG63、121TAPGGNDV128、186MVLDGPHRGGDLRRAR201.鉴定表明,3个B细胞表位有成为牛源无乳链球菌预防性表位疫苗的潜力.
Prediction and Identification of B Cell Epitopes of GapC Protein of Streptococcus agalactiae
To express the Glyceraldehyde-3-Phosphate Dehydrogenase(GapC)protein of Streptococcus agalactiae from bovine and evaluate its immune effect,predict and identify its dominant B cell epitope.In this study,the GapC gene of Streptococcus agalactiae X16087 strain was amplified and the recombinant plasmid pET-28a-WR-X16087 was constructed.After induction and expression,purified GapC protein was used to immunize mice and the immune effect was analyzed.Then bioinformatics analysis was performed to predict and identify the B cell epitopes of GapC protein.The results showed that after induction and purification,the target protein with a size of 44 kDa was obtained.After immunizing,the mice antibody ti-ter reached 1∶218700,and the antibody subtypes mainly were IgG1 and IgG2b;mice protection could reach 100%.GapC immunization could significantly reduce the bacterial load in liver,spleen,lung and kid-ney of mice.GapC protein could reduce the pathological damage caused by bacterial invasion in mice.The B cell epitopes of GapC were predicted and screened.Based on the specific antibodies,three linear B cell epitopes of GapC were identified:46KYDTTQGRFDGTVEVKDG63,121TAPGGNDV128,186MVLDGPHR-GGDLRRAR201.The finding showed that the three B cell epitopes had the potential to be used in prophy-lactic epitope vaccines against bovine mastitis of Streptococcus agalactiae.

Streptococcus agalactiaeGapC proteinimmune effectB cell epitopesscreening and iden-tification

李胜男、王汉青、吕芬芬、赵海利、苏艳

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新疆农业大学 动物医学学院,乌鲁木齐 830052

新疆昭苏县农业农村局,昭苏 835500

无乳链球菌 GapC蛋白 免疫效果 B细胞表位 筛选与鉴定

新疆维吾尔自治区高等学校科研计划重点项目

XJEDU2018I1009

2023

新疆农业大学学报
新疆农业大学

新疆农业大学学报

影响因子:0.599
ISSN:1007-8614
年,卷(期):2023.46(4)
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