已入深夜,您辛苦了!由于当前在线用户较少,发布求助请尽量完整的填写文献信息,科研通机器人24小时在线,伴您度过漫漫科研夜!祝你早点完成任务,早点休息,好梦!

[Cloning, expression of PPMP antigen genes of Clonorchis sinensis and immunogenic identification of the recombinant proteins].

华支睾吸虫 重组DNA 生物 华支睾吸虫病 分子生物学 免疫原性 互补DNA 抗原 基因 cDNA文库 免疫学 遗传学 蠕虫
作者
Xue-Nian Xu,Yan Zhou,Yuting Dong,Tan YuGuang,Yi-Fang Bao,Bin Xu,Na Cheng,Hongbo Xu,Xue‐Ming Li,Zheng Feng
出处
期刊:Chinese Journal of Parasitology and Parasitic Diseases 卷期号:28 (6): 401-405 被引量:1
标识
摘要

Objective To screen and identify new specific antigen gene from a cDNA library of adult Clonorchis sinensis, and investigate the immunogenicity of the recombinant proteins. Methods The lambdaZAP cDNA library of adult C. sinensis was immunoscreened with pooled sera of clonorchiasis patients. The positive clones were sequenced and analyzed. The sequence encoding the mature peptide was cloned into prokaryotic expression vector pET28b(+). The recombinant plasmid was transformed into E. coli BLR21 (DE3) or BLR21 (DE3) pLysS and followed by expression of the protein induced by IPTG. The recombinant protein was purified by His-bind-resin (Ni-NTA) affinity chromatography and identified by Western blotting. BALB/c mice were immunized with purified recombinant pET28b-Cs2 protein, and the sera from immunized mice were analyzed for specific antibodies by ELISA. Results A total of 44 positive clones were isolated from the C. sinensis cDNA library. Three clones containing specific tandem repeats of PPMP amino acid sequence were named as C. sinensis PPMP antigen genes. The genes containing KPPMPGDRDA, QPPMPGGRDA were named as type PPMP I and type PPMP II antigens, respectively. Sequence analysis revealed that these PPMP genes were a novel specific C. sinensis antigen gene family. Two new genes, PPMP I Cs2 and PPMP II Cs3, were expressed in E. coli, and SDS-PAGE showed that the two recombinant proteins were about M(r) 22 000 and M(r) 39 000. The two soluble recombinant proteins were recognized by pooled sera of clonorchiasis patients. A high level of specific IgG against the recombinant proteins (maximum dilution 1 : 64 000) was produced in immunized mice. Conclusion A novel PPMP gene family of C. sinensis has been identified, and its recombinant proteins show high immunogenicity.

科研通智能强力驱动
Strongly Powered by AbleSci AI
更新
大幅提高文件上传限制,最高150M (2024-4-1)

科研通是完全免费的文献互助平台,具备全网最快的应助速度,最高的求助完成率。 对每一个文献求助,科研通都将尽心尽力,给求助人一个满意的交代。
实时播报
研友_VZG7GZ应助青梧采纳,获得10
刚刚
polaris发布了新的文献求助10
刚刚
满意一曲完成签到,获得积分10
1秒前
今后应助thousandlong采纳,获得10
1秒前
上官尔芙发布了新的文献求助10
1秒前
2秒前
李爱国应助ppprotein采纳,获得10
2秒前
高大的机器猫完成签到 ,获得积分10
3秒前
young_joint发布了新的文献求助10
3秒前
发文必中应助BSDL采纳,获得10
4秒前
Nick完成签到,获得积分10
6秒前
7秒前
港岛妹妹完成签到 ,获得积分10
8秒前
倾城完成签到,获得积分10
10秒前
jj完成签到,获得积分10
10秒前
江河发布了新的文献求助10
12秒前
13秒前
莉莉丝完成签到,获得积分10
14秒前
haosu发布了新的文献求助30
14秒前
husgbxud发布了新的文献求助20
14秒前
太懂我了吧完成签到,获得积分10
15秒前
Jiro完成签到,获得积分10
17秒前
17秒前
希望天下0贩的0应助Aurora采纳,获得10
17秒前
18秒前
18秒前
21秒前
义气高丽完成签到 ,获得积分10
21秒前
充电宝应助迷人的Jack采纳,获得10
21秒前
22秒前
SONNG发布了新的文献求助10
22秒前
搜集达人应助wh雨采纳,获得10
24秒前
25秒前
阿洁发布了新的文献求助10
26秒前
27秒前
an发布了新的文献求助10
28秒前
豆子应助guard采纳,获得10
30秒前
31秒前
ding应助Nothing采纳,获得10
35秒前
35秒前
高分求助中
Sustainability in ’Tides Chemistry 2000
Studien zur Ideengeschichte der Gesetzgebung 1000
The ACS Guide to Scholarly Communication 1000
TM 5-855-1(Fundamentals of protective design for conventional weapons) 1000
Handbook of the Mammals of the World – Volume 3: Primates 805
Ethnicities: Media, Health, and Coping 800
Gerard de Lairesse : an artist between stage and studio 500
热门求助领域 (近24小时)
化学 医学 生物 材料科学 工程类 有机化学 生物化学 物理 内科学 纳米技术 计算机科学 化学工程 复合材料 基因 遗传学 催化作用 物理化学 免疫学 量子力学 细胞生物学
热门帖子
关注 科研通微信公众号,转发送积分 3072376
求助须知:如何正确求助?哪些是违规求助? 2726162
关于积分的说明 7492979
捐赠科研通 2373785
什么是DOI,文献DOI怎么找? 1258716
科研通“疑难数据库(出版商)”最低求助积分说明 610362
版权声明 596952