骨桥蛋白腺病毒穿梭质粒的构建及鉴定
onstmetion and identification of recombinant adenoviral shuttle vector with Opn gene
目的 构建腺病毒穿梭质粒pacAd5-opn重组质粒，为探讨构建大鼠骨桥蛋白（OPN）基因的腺病毒载体做好准备，为OPN基因治疗皮肤创口无瘢痕愈合的研究奠定基础。方法：采用RT-PCR方法将OPN基因从大鼠骨组织中克隆出来。将该基因cDNA片段插入腺病毒穿梭质粒pacAd5中，通过分子生物学技术，用PCR、酶切、测序进行鉴定后，转化至高效感受态DH5a细菌细胞中进行扩增，提取质粒，获得重组质粒腺病毒穿梭质粒pacAd5-opn。结果：经分析鉴定，重组质粒中含有OPN基因，达到实验预期目的。结论：成功构建表达OPN基因的重组质粒腺病毒穿梭质粒pacAd5-opn,有助于后续研究构建腺病毒载体Ad-opn。
Objective To construct recombinant adenovirus shuttle plasmid pacAd5-opn. OPN gene was synthesized with reverse transcription polymerase chain reaction (RT-PCR), and then was cloned into adenovirus shuttle plasmid pacAd5. After identification with restriction enzyme analysis, PCR and nucleotide sequencing analysis, and then the recombinant plasmid were transformed into the cells of DH5αbacteria．After amplification and abstraction．the recombinant plasmid pacAd5-opn was obtaind．Results The recombinant plasmid was confirmed containing OPN gene. Conclusion We have constructed recombinant adenovirus shuttle plasmid with OPN successfully. which may be helpful to the next study about the construction of the adenovirus OPN.
王佐林、刘安娜
基础医学生物科学研究方法、生物科学研究技术分子生物学
腺病毒穿梭质粒OPN
adenovirusrecombinant plasmidOPN
王佐林,刘安娜.骨桥蛋白腺病毒穿梭质粒的构建及鉴定[EB/OL].(2012-02-28)[2025-08-16].http://www.paper.edu.cn/releasepaper/content/201202-1080.点此复制
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