{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,4]],"date-time":"2026-06-04T14:03:55Z","timestamp":1780581835631,"version":"3.54.1"},"reference-count":25,"publisher":"Wiley","issue":"9","license":[{"start":{"date-parts":[[2024,9,18]],"date-time":"2024-09-18T00:00:00Z","timestamp":1726617600000},"content-version":"vor","delay-in-days":17,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Immunity Inflam &amp;amp; Disease"],"published-print":{"date-parts":[[2024,9]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:sec><jats:title>Background<\/jats:title><jats:p><jats:italic>Mycoplasma pneumoniae<\/jats:italic> (<jats:italic>MP<\/jats:italic>) 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 <jats:italic>MP<\/jats:italic> inactivated or attenuated vaccine. Therefore, it is necessary to develop a <jats:italic>MP<\/jats:italic> genetic engineering vaccine with influenza virus strain as vector.<\/jats:p><\/jats:sec><jats:sec><jats:title>Methods<\/jats:title><jats:p>In this study, the major antigen genes P1a of <jats:italic>MP<\/jats:italic> adhesion factor P1(3862\u20104554 bases) and P30a of P30(49\u2010822 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\u2010P1a or NS\u2010P30a. The recombinant pHW2000 plasmids containing NS\u2010P1a or NS\u2010P30a were cotransfected with the rest 7 fragments of PR8 into HEK293T cells. After inoculating chicken embryos, the recombinant influenza viruses rFLU\u2010P1a and rFLU\u2010P30a were rescued. RT\u2010PCR and sequencing were used to identify the recombinant viruses. The hemagglutination titers of rFLU\u2010P1a and rFLU\u2010P30a 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.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>P1a or P30a was designed to be inserted into the modified NS gene sequence separately and synthesized successfully. RT\u2010PCR identification of the recombinant viruses rFLU\u2010P1a and rFLU\u2010P30a showed that P1a (693\u2009bp), P30a (774\u2009bp), NS\u2010P1a (1992bp) and NS\u2010P30a (2073\u2009bp) 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.<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusion<\/jats:title><jats:p>The recombinant viruses rFLU\u2010P1a and rFLU\u2010P30a 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\u2010P1a or rFLU\u2010P30a 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 <jats:italic>MP<\/jats:italic> vaccines for human.<\/jats:p><\/jats:sec>","DOI":"10.1002\/iid3.70021","type":"journal-article","created":{"date-parts":[[2024,9,18]],"date-time":"2024-09-18T09:06:58Z","timestamp":1726650418000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["Research of recombinant influenza A virus as a vector for <i>Mycoplasma pneumoniae<\/i> P1a and P30a"],"prefix":"10.1002","volume":"12","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-3701-4994","authenticated-orcid":false,"given":"Liang","family":"Yu","sequence":"first","affiliation":[{"name":"Department of Clinical Laboratory The First People's Hospital of Yunnan Province Kunming China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Wang","family":"Yongbo","sequence":"additional","affiliation":[{"name":"Department of Clinical Laboratory The First People's 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Medical College Kunming China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ma","family":"Linna","sequence":"additional","affiliation":[{"name":"Department of Medical Laboratory Technique Kunming Medical University Haiyuan College Kunming China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2024,9,18]]},"reference":[{"key":"e_1_2_8_2_1","doi-asserted-by":"publisher","DOI":"10.3390\/vaccines11020469"},{"issue":"3","key":"e_1_2_8_3_1","first-page":"169","article-title":"June","volume":"34","author":"G\u00f3mez Rufo D","year":"2021","journal-title":"Rev Esp 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