| NT4-SAC-HA2-TAT融合基因表达载体的构建及鉴定 |
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| 引用本文： | 张士宝,刘庆勇,阮喜云,陈杰,张建军,李宗武,杨广笑,王全颖.NT4-SAC-HA2-TAT融合基因表达载体的构建及鉴定[J].山东大学学报(医学版),2009,47(6):15-19. |
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| 作者姓名： | 张士宝 刘庆勇 阮喜云 陈杰 张建军 李宗武 杨广笑 王全颖 |
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| 作者单位： | 张士宝,刘庆勇,陈杰,张建军,李宗武,ZHANG Shi-bao,LIU Qing-yong,CHEN Jie,ZHANG Jian-jun,LI Zong-wu(山东大学附属济南市中心医院泌尿外科,济南,250013);阮喜云,RUAN Xi-yun(山东大学附属济南市中心医院神经内科,济南,250013);杨广笑,王全颖,YANG Guang-xiao,WANG Quan-ying(西安华广生物工程公司,西安,710025)
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| 摘 要： | 目的 构建NT4-SAC-HA2-TAT融合肽cDNA克隆，探讨前列腺凋亡反应基因-4(Par-4)核心区凋亡肽(SAC)作为目的基因对前列腺癌的治疗作用。 方法 应用互为引物模板法合成两端具有NaeI和KpnI酶识别位点的SAC cDNA片段。将所得SAC和已有HA2-TAT片段克隆到具有相应酶切位点的pBV220-NT4质粒，得到pBV220-NT4-SAC-HA2-TAT重组质粒。PCR扩增NT4-SAC-HA2-TATcDNA片段，并将其克隆到pGEM-T easy中，转化细菌筛选阳性克隆，酶切鉴定，序列测定分析。 结果 经DNA测序、限制性内切酶酶切等证实，PCR获得了编码NaeI和KpnI酶切位点的SAC cDNA片段，并将NT4、SAC、HA2-TAT亚克隆于pBV220内。 结论 成功构建了NT4-SAC-HA2-TAT融合基因的原核表达载体pBV220-NT4-SAC-HA2-TAT。
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| 关 键 词： | 前列腺凋亡反应基因-4 融合肽 前列腺肿瘤 基因疗法 |
| 收稿时间： | 2008-12-04 |
onstruction and identification of the expression vector ofNT4-SAC-HA2-TAT fusion gene |
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| ZHANG Shi-Bao,LIU Qing-Yong,RUAN Xi-Yun,CHEN Jie,ZHANG Jian-Jun,LI Zong-Wu,YANG Guang-Xiao,WANG Quan-Ying.onstruction and identification of the expression vector ofNT4-SAC-HA2-TAT fusion gene[J].Journal of Shandong University:Health Sciences,2009,47(6):15-19. |
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| Authors: | ZHANG Shi-Bao LIU Qing-Yong RUAN Xi-Yun CHEN Jie ZHANG Jian-Jun LI Zong-Wu YANG Guang-Xiao WANG Quan-Ying |
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| Institution: | 1.Department of Urinary Surgery;Jinan Central Hospital Affiliated to Shandong University;Jinan 250013;China;2.Department of Neurology;Jinan Central Hospital Affiliated to ShandongUniversity;3.Xi'an Huaguang Biological Engineering Co.Ltd;Xi'an 710025;China |
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| Abstract: | Objective To construct the prokaryocyte expression vector bearing fusion gene NT4-SAC-HA2-TAT and lay a foundation for further study on the genetic therapy of prostate cancer.Methods By means of asymmetrical primer/template,the fragment encoding SAC was gained by the PCR method.Then the SAC/NaeI,KpnI and HA2-TAT/KpnI,and Xho I were cloned into recombinant vector pBV220-NT4/NaeI,SalI and the recombinant plasmid pBV220-NT4-SAC-HA2-TAT was obtained.The fragment NT4SAC-HA2-TAT obtained was amplified by PCR.Then... |
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| Keywords: | Par-4 fusion peptide Prostate neoplasms Gene therapy |
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