發(fā)布時(shí)間：2018-03-23 15:58

  本文選題：hIL-　切入點(diǎn)：重組腺病毒載體　出處：24的構(gòu)建及病毒滴度測(cè)定

【摘要】：目的構(gòu)建重組腺病毒載體pDC316-hIL-24,并測(cè)定病毒滴度。方法從HEK293細(xì)胞中提取mRNA,設(shè)計(jì)特異性引物,通過RT-PCR法擴(kuò)增hIL-24基因全編碼區(qū)序列。將測(cè)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(gòu)建重組穿梭質(zhì)粒pDC316-EGFP-hIL-24,在脂質(zhì)體介導(dǎo)下與腺病毒輔助大質(zhì)粒pBHGlox(delta)E1,3Cre共轉(zhuǎn)染293細(xì)胞,包裝產(chǎn)生復(fù)制缺陷型重組腺病毒pDC316-hIL-24,經(jīng)HEK293細(xì)胞擴(kuò)增,TCID50法測(cè)定重組腺病毒滴度。結(jié)果成功構(gòu)建出表達(dá)h IL-24基因的重組腺病毒載體(pDC316-hIL-24),獲得了高滴度表達(dá)hIL-24基因的重組腺病毒。結(jié)論重組腺病毒表達(dá)載體(pDC315-hIL-24)的成功構(gòu)建及重組腺病毒的獲得有利于進(jìn)一步開展腫瘤的轉(zhuǎn)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(hào)：1654149