高级检索
当前位置: 首页 > 详情页

Research of recombinant influenza A virus as a vector for Mycoplasma pneumoniae P1a and P30a

文献详情

资源类型:
WOS体系:

收录情况: ◇ SCIE

机构: [1]First Peoples Hosp Yunnan Prov, Dept Clin Lab, Kunming, Peoples R China [2]Chinese Acad Med Sci & Peking Union Med Coll, Inst Med Biol, Dept Biol Prod 5, 935 Jiaoling Rd, Kunming 650031, Peoples R China [3]Kunming Med Univ, Haiyuan Coll, Dept Med Lab Tech, 389 Haiyuan North Rd, Kunming 650106, Yunnan, Peoples R China
出处:
ISSN:

关键词: genetic engineering influenza virus vector Mycoplasma pneumoniae vaccine

摘要:
Background: Mycoplasma pneumoniae (MP) MP ) is a common respiratory pathogen affecting the longevity of the elderly and the health of children. However, the human vaccine against MP has not been successfully developed till now due to the poor immunogenicity and side effects of MP inactivated or attenuated vaccine. Therefore, it is necessary to develop a MP genetic engineering vaccine with influenza virus strain as vector. Methods: In this study, the major antigen genes P1a of MP adhesion factor P1(3862-4554 - 4554 bases) and P30a of P30(49-822 - 822 bases) were inserted into the nonstructural protein (NS) gene of Influenza A virus strain A/Puerto Rio/8/ 34(H1N1), PR8 for short, to construct the recombinant vectors NS-P1a - P1a or NS-P30a. - P30a. The recombinant pHW2000 plasmids containing NS-P1a - P1a or NS-P30a - P30a were cotransfected with the rest 7 fragments of PR8 into HEK293T cells. After inoculating chicken embryos, the recombinant influenza viruses rFLU-P1a - P1a and rFLU-P30a - P30a were rescued. RT-PCR - PCR and sequencing were used to identify the recombinant viruses. The hemagglutination titers of rFLU-P1a - P1a and rFLU- - P30a were determined after five successive generations in chicken embryos so as to indicate the genetic stability of the recombinant viruses. The morphology of recombinant influenza viruses was observed under electron microscopy. Results: P1a or P30a was designed to be inserted into the modified NS gene sequence separately and synthesized successfully. RT-PCR - PCR identification of the recombinant viruses rFLU-P1a - P1a and rFLU-P30a - P30a showed that P1a (693 bp), P30a (774 bp), NS-P1a - P1a (1992bp) and NS-P30a - P30a (2073 bp) bands were found, and the sequencing results were correct. After five successive generations, each virus generation has a certain hemagglutination titer (from 1:32 to 1:64), and the band of P1a or P30a can be seen in the corresponding positions. The virus particles under the electron microscope appeared as spheres or long strips connected by several particles, revealing a complete viral membrane structure composed of virus lipid bilayer, hemagglutinin, neuraminidase, and matrix proteins. Conclusion: The recombinant viruses rFLU-P1a - P1a and rFLU-P30a - P30a which carried the advantaged immune regions of the P1 and P30 genes in MP were successfully constructed and identified. And the genetic stability of rFLU-P1a - P1a or rFLU-P30a - P30a was relatively high. The typical and complete morphology of influenza virus was observed under the electron microscope. Our research provided a foundation for the further development of MP vaccines for human.

基金:
语种:
WOS:
PubmedID:
中科院(CAS)分区:
出版当年[2024]版:
最新[2023]版:
大类 | 4 区 医学
小类 | 4 区 免疫学
JCR分区:
出版当年[2023]版:
Q3 IMMUNOLOGY
最新[2023]版:
Q3 IMMUNOLOGY

影响因子: 最新[2023版] 最新五年平均 出版当年[2023版] 出版当年五年平均 出版前一年[2022版]

第一作者:
第一作者机构: [1]First Peoples Hosp Yunnan Prov, Dept Clin Lab, Kunming, Peoples R China
通讯作者:
推荐引用方式(GB/T 7714):
APA:
MLA:

资源点击量:83046 今日访问量:0 总访问量:682 更新日期:2025-01-01 建议使用谷歌、火狐浏览器 常见问题

版权所有©2020 云南省第一人民医院 技术支持:重庆聚合科技有限公司 地址:云南省昆明市西山区金碧路157号