[1]李建成,彭世清,胡章立.SEC2在莱茵衣藻中的表达及免疫学活性分析[J].深圳大学学报理工版,2012,29(No.2(095-188)):159-164.[doi:10.3724/SP.J.1249.2012.02159]
 LI Jian-cheng,PENG Shi-qing,et al.Expression and immunological activity of enterotoxin C2 from Staphylococcus aureus in Chlamydomonas reinhardti[J].Journal of Shenzhen University Science and Engineering,2012,29(No.2(095-188)):159-164.[doi:10.3724/SP.J.1249.2012.02159]
点击复制

SEC2在莱茵衣藻中的表达及免疫学活性分析()
分享到:

《深圳大学学报理工版》[ISSN:1000-2618/CN:44-1401/N]

卷:
第29卷
期数:
2012年No.2(095-188)
页码:
159-164
栏目:
生物工程
出版日期:
2012-03-23

文章信息/Info

Title:
Expression and immunological activity of enterotoxin C2 from Staphylococcus aureus in Chlamydomonas reinhardti
作者:
李建成13 彭世清3 胡章立2
1)海南大学农学院,海口 570228
2)深圳大学生命科学学院,深圳市海洋生物资源与生态环境重点实验室, 深圳 518060
3)中国热带农业科学院热带生物技术研究所,农业部热带作物生物技术重点开放实验室,海口 571101
Author(s):
LI Jian-cheng1 3 PENG Shi-qing3 and HU Zhang-li2
1) College of Agriculture, Hainan University, Haikou 570228, P.R.China
2) Shenzhen Key Laboratory of Marine Bioresource and Eco-environmental Science, College of Life Science, Shenzhen University, Shenzhen 518060, P.R.China
3) Ministry of Agriculture Key Biotechnology Laboratory for Tropical Crops, The Institute of Tropical Bioscience and Biotechnology, Chinese Academy of Tropical Agricultural Sciences, Haikou 571101, P.R.China
关键词:
基因工程金黄色葡萄球菌肠毒素C2莱茵衣藻超抗原转基因藻抗肿瘤定点突变
Keywords:
genetic engineering Staphylococcus aureus enterotoxin C2 Chlamydomonas reinhardtii superantigen transgenic algae anti-tumor site-directed mutagenesis
分类号:
Q 37;R 392.1
DOI:
10.3724/SP.J.1249.2012.02159
文献标志码:
A
摘要:
通过将金黄色葡萄球菌(Staphylococcus aureus, S.aureus)肠毒素C2(staphylococcal enterotoxin C2,SEC2)基因进行定点突变,获得具有超抗原性的减毒C2基因序列sec2t,把该基因插入含Hsp70A-RBCS2启动子和RBCS2终止子的pH124载体上,构建莱茵衣藻表达载体pH124sec2t,采用“珠磨法”将该载体转入细胞壁缺陷型莱茵衣藻(Chlamydomonas reinhardtii)CC-849中,经含10 μg/mL Zeomycin的抗性平板筛选、PCR及RT-PCR分析,获得转基因莱茵衣藻Tran-sec2t.Southern blot和Western blot分析表明,sec2t基因已整合入莱茵衣藻核基因组中,且能表达相对分子质量为26 kDa(1 Da=1 u)的目的蛋白.分别以CC-849和Tran-sec2t两种藻细胞培养液(每毫升106个细胞)喂食Balb/c小鼠,并用流式细胞仪检测分析小鼠CD3+、CD4+和CD8+细胞,结果发现,可表达减毒超抗原SEC2T的转基因莱茵衣藻Tran-sec2t能刺激小鼠T淋巴细胞的产生;与对照相比,CD4+与CD8+细胞数目比值有所降低,但CD3+和CD8+细胞有明显提高.研究结果有助于利用转基因藻生产抗肿瘤制剂和动物免疫增强剂.
Abstract:
A superantigenic gene(sec2t) of the attenuated enterotoxin C2 was obtained by the site-directed mutagenesis of enterotoxin C2 in Staphylococcus aureus. The sec2t was inserted into the pH124 vector with Hsp70A-RBCS2 promoter and RBCS2 terminator to construct the expression vectors pH124sec2t for transformation in Chlamydomonas reinhardtii. The pH124sec2t vectors were transformed into the algal cell by the glass-bead method in Chlamydomonas reinhardtii CC-849. A number of transformants appeared in the TAP plates with 10 μg/mL of Zeomycin in 3~4 weeks. The transgenic strain Tran-sec2t was further screened by PCR and RT-PCR
analysis. Southern blot and Western blot analysis showed that sec2t gene had been integrated into the nuclear genome of Tran-sec2t, and a 26 kDa(1 Da=1 u) SEC22T protein had been expressed in the Tran-sec2t. In order to detect the immunological activity of Tran-sec2t, the saline (0.9%) and cultures (106 / mL) of CC-849 and Tran-sec2t were used to feed the three groups of Balb/c mice respectively. The CD3+, CD4+ and CD8+ of experimental mice were detected by flow cytometry. The results show that T lymphocytes proliferation of mice is stimulated by the SEC2T expressed in Tran-sec2t. To compare with T-cell response of CC-849, the Tran-sec2t leads to the increase of CD3+ and CD8+ significantly, but it decreases the ratio of CD4+/CD8+ in the experimental mice.

参考文献/References:

[1] Baker M D, Acharya K R. Superantigens: structure-function relationships [J]. International Journal of Medical Microbiology, 2004, 293(7/8): 529-537.
[2] XIA Zhan-qiang. Research progress for the superantigen enterotoxin of Staphylococcus aureus [J]. Chinese Practical Medicine, 2009, 31(4): 235-236.(in Chinese)
夏占强. 超抗原SE-金葡菌肠毒素的研究进展[J]. 中国实用医药, 2009, 31(4): 235-236.
[3] Baker M D, Papageorgious A C. Titball R W, et al. Structural and functioml role of threonine 112 in a super-antigen Staphylococcal aureus enterotoxin B[J]. The Journal of Biological Chemistry, 2002, 277(4): 2756-2762.
[4] Liewelyn M,Cohen J. Superantigens: microbial agents that corrupt immunity[J]. The Lancet Infectious Disease, 2002, 2(3): 156-162.


[5] Wright A, Anderws P L,Titball R N. Induction of emetic, pyreic and behacioral effects of Staphylococcus aureus enterotoxin B in the ferret[J]. Infection and Immunity, 2000, 68(4): 2386-2389.
[6] HU Zhang-li, ZHAO Zhong-lin, WU Zhi-hua, et al. Successful expression of heterologous egfp gene in the mitochondria of a photosynthetic eukaryote Chlamydomonas reinhardtii [J]. Mitochondrion, 2011, 11(5): 716-721.
[7] Mayfield S P, Franklin S, Lerner R. Expression and assembly of a fully active antibody in algae [J]. Proceedings of the National Academy of Sciences of the United States of America, 2003, 100(2): 438-442.
[8] SUN Meng, QIAN Kai-xian, SU Ning, et al. Foot-and-mouth disease virus VP1 protein fused with cholera toxin B subunit expressed in Chlamydomonas reinhardtii chloroplast [J]. Biotechnology, 2003, 25(13): 1087-1092.
[9] WANG Chao-gang, HU Zhang-li, LEI An-ping, et al. Biosynthesis of poly-3-hydroxybutyrate (PHB) in transgenic green algae Chlamydomonas reinhardtii[J]. Journal of Phycology, 2010, 46(2): 396-402.
[10] Schlievert P M, Shands K N, Dan B B, et al. Role of super-antignes in human disease[J]. The Journal of Infectious Diseases, 1993, 167(5): 997.
[11] Madeleine W Cunningham. T cell mimicry in inflammatory heart disease[J]. Molecular Immunology, 2004, 40(14/15): 1121-1127.
[12] Krakauer T, Chen X, Howard O M, et al. Triptolide attenuates endotoxin and staphylococcal exotoxin-induced T-cell proliferation and production of cytokines and chemokines [J]. Immunopharmacol. Immunotoxicol, 2005, 27(1): 53-66.

相似文献/References:

[1]龚映雪,台艳,肖文娟,等.酵母多基因表达载体在纤维素生物转化中应用[J].深圳大学学报理工版,2010,27(1):82.
 GONG Ying-xue,TAI Yan,XIAO Wen-juan,et al.Construction of Saccharomyces cerevisiae integrated expression vector and its application in cellulose bioconvesion[J].Journal of Shenzhen University Science and Engineering,2010,27(No.2(095-188)):82.
[2]莫蓓莘.RNA在植物细胞中的定位及其研究进展[J].深圳大学学报理工版,2011,28(No.3(189-282)):230.
 MO Bei-xin.RNA localization in plant cells and advances of the research[J].Journal of Shenzhen University Science and Engineering,2011,28(No.2(095-188)):230.
[3]刘士德,陈汗英,张建华,等.PSTI1可提高大肠杆菌应答胁迫能力[J].深圳大学学报理工版,2011,28(No.4(283-376)):347.
 LIU Shi-de,CHEN Han-ying,ZHANG Jian-hua,et al.A novel STI1-like protein from Physarum polycephalum can enhance E.coli response to stress[J].Journal of Shenzhen University Science and Engineering,2011,28(No.2(095-188)):347.
[4]张毅哲,刘琼,田静,等.人脑中脱碘酶相互作用蛋白的筛选与验证[J].深圳大学学报理工版,2011,28(No.5(377-470)):423.
 ZHANG Yi-zhe,LIU Qiong,TIAN Jing,et al.Investigation on the interactive protein of iodothyronine deiodinase 3 mutant in human brain[J].Journal of Shenzhen University Science and Engineering,2011,28(No.2(095-188)):423.
[5]王潮岗,黄惠珠,孙海珊,等.一种莱茵衣藻启动子功能检测系统的构建[J].深圳大学学报理工版,2012,29(No.6(471-580)):534.[doi:10.3724/SP.J.1249.2012.06534]
 WANG Chao-gang,HUANG Hui-zhu,SUN Hai-shan,et al.Construction of Chlamydomonas reinhardtii system for analyzing the function of algal promoter[J].Journal of Shenzhen University Science and Engineering,2012,29(No.2(095-188)):534.[doi:10.3724/SP.J.1249.2012.06534]
[6]刘泽寰,郑阳阳,龚映雪,等.利用重组酵母实现木薯至乙醇的直接转化[J].深圳大学学报理工版,2012,29(No.6(471-580)):548.[doi:10.3724/SP.J.1249.2012.06548]
 LIU Ze-huan,ZHENG Yang-yang,GONG Ying-xue,et al.Direct conversion from cassava to bioethanol with a recombinant strain of yeast[J].Journal of Shenzhen University Science and Engineering,2012,29(No.2(095-188)):548.[doi:10.3724/SP.J.1249.2012.06548]
[7]胡章立,李建成,程雪薇.莱茵衣藻线粒体遗传系统及其外源基因表达[J].深圳大学学报理工版,2013,30(No.6(551-660)):603.[doi:10.3724/SP.J.1249.2013.06603]
 Hu Zhangli,Li Jiancheng,and Cheng Xuewei.The mitochondrial genetic system and exogenous gene expression of Chlamydomonas reinhardtii[J].Journal of Shenzhen University Science and Engineering,2013,30(No.2(095-188)):603.[doi:10.3724/SP.J.1249.2013.06603]
[8]王潮岗,李逸,胡章立.转lpaat和gpd1基因工程藻的快速筛选[J].深圳大学学报理工版,2016,33(No.1(001-110)):33.[doi:10.3724/SP.J.1249.2016.01033]
 Wang Chaogang,Li Yi,and Hu Zhangli.Rapid screening of the transgenic algae haboring lpaat and gpd1[J].Journal of Shenzhen University Science and Engineering,2016,33(No.2(095-188)):33.[doi:10.3724/SP.J.1249.2016.01033]
[9]邓旭,黄慧炜,梁彩柳,等.一株海洋细菌高汞抗性及高富集能力的分析[J].深圳大学学报理工版,2016,33(No.6(551-660)):551.[doi:10.3724/SP.J.1249.2016.06551]
 Deng Xu,Huang Huiwei,Liang Cailiu,et al.Analysis of a marine bacterium with high mercury resistance and bioaccumulation capacity[J].Journal of Shenzhen University Science and Engineering,2016,33(No.2(095-188)):551.[doi:10.3724/SP.J.1249.2016.06551]
[10]凌文康,黄慧炜,刘冰,等.贻贝足丝蛋白的汞、镉抗性及富集机制[J].深圳大学学报理工版,2019,36(No.4(347-472)):361.[doi:10.3724/SP.J.1249.2019.04361]
 LING Wenkang,HUANG Huiwei,LIU Bing,et al.Mercury and cadmium resistance and enrichment mechanism of mussel foot fibroin[J].Journal of Shenzhen University Science and Engineering,2019,36(No.2(095-188)):361.[doi:10.3724/SP.J.1249.2019.04361]

备注/Memo

备注/Memo:
Received:2012-01-04;Accepted:2012-02-14
Foundation:NationalNaturalScienceFoundationofChina(31070323),ScienceandTechnologyPlanningProjectofGuangdongProvince(2009B020307013)
Corresponding author:Professor HU Zhang-li. E-mail: huzl@szu.edu.cn
Citation:LI Jian-cheng, PENG Shi-qing, HU Zhang-li. Expression and immunological activity of enterotoxin C2 from Staphylococcus aureus in Chlamydomonas reinhardtii[J]. Journal of Shenzhen University Science and Engineering, 2012, 29(2): 159-164.(in Chinese)
基金项目:国家自然科学基金资助项目(31070323) ,广东省科技计划资助项目(2009B020307013)
作者简介:李建成(1985-),男 (汉族),湖南省常德市人,海南大学硕士研究生. E-mail: lijc1007@163.com
引文:李建成,彭世清,胡章立. SEC2在莱茵衣藻中表达及免疫学活性分析[J]. 深圳大学学报理工版,2012,29(2):159-164.
更新日期/Last Update: 2012-03-28