[1]惠萌萌孙曼曼,刘秀霞,杨艳坤,等.猪伪狂犬病病毒糖蛋白gD在谷氨酸棒杆菌中的表达及优化[J].江苏农业学报,2022,38(06):1578-1585.[doi:doi:10.3969/j.issn.1000-4440.2022.06.016]
　HUI Meng-meng,SUN Man-man,LIU Xiu-xia,et al.Expression and optimization of glycoprotein gD of porcine pseudorabies virus in Corynebacterium glutamicum[J].,2022,38(06):1578-1585.[doi:doi:10.3969/j.issn.1000-4440.2022.06.016]
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猪伪狂犬病病毒糖蛋白gD在谷氨酸棒杆菌中的表达及优化()
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江苏农业学报[ISSN:1006-6977/CN:61-1281/TN]

卷:
38
期数:
2022年06期
页码:
1578-1585
栏目:
畜牧兽医·水产养殖
出版日期:
2022-12-31

文章信息/Info

Title:
Expression and optimization of glycoprotein gD of porcine pseudorabies virus in Corynebacterium glutamicum
作者:
惠萌萌1孙曼曼1; 刘秀霞1; 2; 3; 杨艳坤1; 2; 3; 白仲虎1; 2; 3
(1.江南大学粮食发酵与食品生物制造国家工程研究中心，江苏无锡214122；2.江南大学工业生物技术教育部重点实验室，江苏无锡214122；3.江苏省生物活性制品加工工程技术研究中心，江苏无锡214122)
Author(s):
HUI Meng-meng1; SUN Man-man1; LIU Xiu-xia1; 2; 3; YANG Yan-kun1; 2; 3; BAI Zhong-hu1; 2; 3
(1.National Engineering Research Center of Cereal Fermentation and Food Biomanufacturing, Jiangnan University, Wuxi 214122, China;2.Key Laboratory of Industrial Biotechnology, Ministry of Education, Jiangnan University, Wuxi 214122, China;3.Jiangsu Provincial Engineering Research Center for Bioactive Product Processing, Wuxi 214122, China)
关键词:
谷氨酸棒杆菌; 猪伪狂犬病病毒; 糖蛋白gD; 发酵优化
Keywords:
Corynebacterium glutamicum; porcine pseudorabies virus; glycoprotein gD; fermentation optimization
分类号:
S858.282.65+9.1
DOI:
doi:10.3969/j.issn.1000-4440.2022.06.016
文献标志码:
A
摘要:
为在谷氨酸棒杆菌（Corynebacterium glutamicum）中进行猪伪狂犬病病毒糖蛋白gD（Glycoprotein gD of porcine pseudorabies virus）的可溶性表达，通过启动子、底盘菌株和发酵条件的优化，提高gD在C. glutamicum中的表达量，使用十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（SDS-PAGE）和Western Blot方法进行表达产物的分析与鉴定，用镍离子亲和层析柱纯化产物。SDS-PAGE和Western Blot分析结果表明，gD在谷氨酸棒杆菌表达系统获得正确表达，相对分子质量约4.5×104，可与猪抗gD阳性血清特异性反应。摇瓶培养下最优发酵条件为诱导温度30 ℃，诱导时长24 h，优化后gD质量浓度可达到517 mg/L。
Abstract:
To express soluble glycoprotein gD of porcine pseudorabies virus in Corynebacterium glutamicum, promoter, chassis strain and fermentation conditions were optimized in this study to improve the expression of gD in C. glutamicum. Then the target protein gD was detected by sodium dodecyl subfate-polyacrylamide gel electrophoresis(SDS-PAGE) and Western Blot, and was purified by Ni-NTA affinity chromatography column. The results showed that glycoprotein gD was successfully expressed in C. glutamicum with a relative molecular mass of about 4.5×104, and the protein could specifically bind to PRV antibodies. Under the optimal shaking flask culture condition, the gD production reached the highest mass concentration (517 mg/L) after 24 h incubation at 30 ℃.

参考文献/References:

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备注/Memo

备注/Memo:
收稿日期：2022-02-25基金项目：国家自然科学基金项目（22078128、21878124）作者简介：惠萌萌（1996-），女，安徽宿州人，硕士，主要从事发酵工程、医药蛋白表达等方面的研究。（E-mail）huimengmengw@163.com通讯作者：白仲虎，（E-mail）baizhonghu@jiangnan.edu.cn
更新日期/Last Update: 2023-01-13