發(fā)布時間：2018-03-23 15:58

  本文選題：hIL-　切入點：重組腺病毒載體　出處：24的構建及病毒滴度測定

【摘要】：目的構建重組腺病毒載體pDC316-hIL-24,并測定病毒滴度。方法從HEK293細胞中提取mRNA,設計特異性引物,通過RT-PCR法擴增hIL-24基因全編碼區(qū)序列。將測序正確的片段用BglⅡ和HindⅢ雙酶切定向插入到pDC316-EGFP穿梭質(zhì)粒中。構建重組穿梭質(zhì)粒pDC316-EGFP-hIL-24,在脂質(zhì)體介導下與腺病毒輔助大質(zhì)粒pBHGlox(delta)E1,3Cre共轉染293細胞,包裝產(chǎn)生復制缺陷型重組腺病毒pDC316-hIL-24,經(jīng)HEK293細胞擴增,TCID50法測定重組腺病毒滴度。結果成功構建出表達h IL-24基因的重組腺病毒載體(pDC316-hIL-24),獲得了高滴度表達hIL-24基因的重組腺病毒。結論重組腺病毒表達載體(pDC315-hIL-24)的成功構建及重組腺病毒的獲得有利于進一步開展腫瘤的轉基因治療研究。
[Abstract]:Objective to construct recombinant adenovirus vector pDC316-hIL-24 and determine its titer. Methods mRNAs were extracted from HEK293 cells and specific primers were designed. The whole coding region of hIL-24 gene was amplified by RT-PCR. The correct sequence was inserted into pDC316-EGFP shuttle plasmid by double enzyme digestion of Bgl 鈪，

本文編號：1654149