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发明授权
CN106011087B S1基因和TM-1基因重组腺病毒的构建方法及重组腺病毒和应用 有权

S1基因和TM-1基因重组腺病毒的构建方法及重组腺病毒和应用
摘要：
本发明公开了一种S1基因和TM‑1基因重组腺病毒的构建方法及重组腺病毒和应用，从IBV H52株和MG S6株中分别扩增出S1基因和TM‑1基因，将获得的目的基因克隆到穿梭载体pDC315‑MCS‑EGFP中，利用脂质体转染法将重组腺病毒穿梭质粒pDC315‑S1‑TM‑1‑EGFP与腺病毒大骨架pBHGlox(delta)E1,3Cre共转染HEK293细胞，重组并包装获得含有S1基因和TM‑1基因的重组腺病毒pBH‑S1‑TM‑1‑EGFP；本发明用于免疫鸡以预防鸡传染性支气管炎和慢性呼吸道病，将此重组腺病毒免疫鸡后，鸡的抗体水平得到显著提高，且对鸡产生有效的保护作用。
摘要（英）：

The invention discloses a construction method of S1 gene and TM-1 gene recombinant adenovirus as well as recombinant adenovirus and application. The construction method comprises the following steps: amplifying a IBV H52 strain and a MG S6 strain to obtain S1 gene and TM-1 gene respectively; cloning the obtained target gene into a shuttle vector pDC315-MCS-EGFP; co-transfecting HEK293cell with the recombinant adenovirus shuttle plasmid pDC315-S1-TM-1-EGFP and adenovirus large skeleton pBHGlox(delta)E1, 3Cre by a liposome transfection method; and recombining and packaging to obtain recombinant adenovirus pBH-S1-TM-1-EGFP containing the S1 gene and TM-1 gene. The recombinant adenovirus disclosed by the invention is used for immunizing chicken to prevent infectious bronchitis and chronic respiratory disease; and by adopting the recombinant adenovirus to immunize chicken, the antibody level of chicken is remarkably raised, and an effective protection effect on chicken is realized.

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