CN101532031B - 非整合慢病毒载体系统及其制备与应用 - Google Patents
非整合慢病毒载体系统及其制备与应用 Download PDFInfo
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- CN101532031B CN101532031B CN2009100469456A CN200910046945A CN101532031B CN 101532031 B CN101532031 B CN 101532031B CN 2009100469456 A CN2009100469456 A CN 2009100469456A CN 200910046945 A CN200910046945 A CN 200910046945A CN 101532031 B CN101532031 B CN 101532031B
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Abstract
Description
纯化的DNA质粒(1ug/μl) | 2μl |
10×buffer | 5μl |
100×BSA | 0.5μl |
Swa I(10U/μl) | 1μl |
Sac I(10U/μl) | 1μl |
H2O | 40.5μl |
Total | 50μl |
试剂 | 体积(ul) |
Primer(+) | 0.4 |
Primer(-) | 0.4 |
ddH2O | 14.7 |
10×buffer | 2 |
MgCl2 | 0.5 |
DNTPs(2.5mM) | 0.8 |
pfu polymerase | 0.2 |
Template(10ng/μl) | 1 |
PCR产物(100ng/μl) | 5μl |
10×buffer | 5μl |
100×BSA | 0.5μl |
Swa I(10U/μl) | 1μl |
Sac I(10U/μl) | 1μl |
H2O | 40.5μl |
Total | 50μl |
试剂 | 对照1(μl) | 对照2(μl) | 连接组1(μl) |
双酶切回收的载体DNA 100ng/μl | 1 | - | 1 |
双酶切回收的PCR产物100ng/μl | - | 1 | 1 |
10×T4噬菌体DNA连接酶缓冲液 | 1 | 1 | 1 |
T4噬菌体DNA连接酶 | 1 | 1 | 1 |
dd H2O | 7 | 7 | 7 |
Total | 10 | 10 | 10 |
Claims (7)
Priority Applications (1)
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CN2009100469456A CN101532031B (zh) | 2009-03-03 | 2009-03-03 | 非整合慢病毒载体系统及其制备与应用 |
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CN2009100469456A CN101532031B (zh) | 2009-03-03 | 2009-03-03 | 非整合慢病毒载体系统及其制备与应用 |
Publications (2)
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CN101532031A CN101532031A (zh) | 2009-09-16 |
CN101532031B true CN101532031B (zh) | 2011-04-20 |
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CN2009100469456A Active CN101532031B (zh) | 2009-03-03 | 2009-03-03 | 非整合慢病毒载体系统及其制备与应用 |
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Families Citing this family (6)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN102199616B (zh) * | 2011-03-07 | 2013-04-24 | 首都医科大学附属北京佑安医院 | Hiv-1表型耐药检测载体及其构建方法 |
CN102229963B (zh) * | 2011-05-11 | 2013-02-13 | 浙江省农业科学院 | 一种慢病毒载体系统及其制备方法 |
CN103540616B (zh) * | 2013-10-23 | 2015-08-12 | 中国人民解放军南京军区福州总医院 | 表达白喉毒素a片段的慢病毒载体系统及其制备与应用 |
CN104910278B (zh) * | 2015-05-27 | 2018-05-25 | 上海吉凯基因科技有限公司 | 一种用于制备cart细胞的具有高效转染能力和生物学活性的慢病毒 |
CN104892770B (zh) * | 2015-05-27 | 2018-05-25 | 上海吉凯基因科技有限公司 | 一种对t细胞和造血干细胞具有高效感染和促增殖能力的慢病毒载体 |
CN107043785B (zh) * | 2016-02-05 | 2021-04-27 | 上海吉凯基因医学科技股份有限公司 | 一种整合型慢病毒载体表达系统 |
Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN1261404A (zh) * | 1997-05-13 | 2000-07-26 | 北卡罗来纳-查佩尔山大学 | 基于慢病毒的基因转移载体 |
WO2002097104A1 (fr) * | 2001-06-01 | 2002-12-05 | Centre National De La Recherche Scientifique | Pseudotypage des vecteurs vih-1 par des enveloppes de rhabdovirus |
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2009
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Patent Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN1261404A (zh) * | 1997-05-13 | 2000-07-26 | 北卡罗来纳-查佩尔山大学 | 基于慢病毒的基因转移载体 |
WO2002097104A1 (fr) * | 2001-06-01 | 2002-12-05 | Centre National De La Recherche Scientifique | Pseudotypage des vecteurs vih-1 par des enveloppes de rhabdovirus |
Non-Patent Citations (2)
Title |
---|
LUIGI NALDINI等.Efficient transfer, integration, and sustained long-term expression of the transgene in adult rat brains injected with a lentiviral vector.《Proc. Natl. Acad. Sci.》.1996,第93卷第11382-11388页. * |
Luigi Naldini等.In Vivo Gene Delivery and Stable Transduction of Nondividing Cells by a Lentiviral Vector.《SCIENCE》.1996,第272卷263-267. * |
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CN101532031A (zh) | 2009-09-16 |
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