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报告基因质粒

pGL6-TA (报告基因质粒)
产品编号: D2105-1μg
产品包装: 1μg
产品价格: 888.00元
产品简介
使用说明
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产品编号 产品名称 产品包装 产品价格
D2105-1μg pGL6-TA (报告基因质粒) 1μg 888.00元

      pGL6-TA (报告基因质粒)是碧云天自行研发的用于在哺乳动物细胞中进行萤火虫萤光素酶(firefly luciferase, 也称荧光素酶)报告基因检测的新一代质粒。该报告基因质粒比Promega公司的pGL3系列有了全面的改进,一方面对于luciferase的编码进行了改进,确保能更好地在哺乳动物细胞中进行表达,同时对整个质粒中所有可以被预测出的可能的转录因子结合位点全部进行了适当的突变处理,在保持原有功能不变的情况下,使各种转录因子在质粒上的非特异性结合降到最低。
      萤光素、萤光素酶、萤火虫萤光素酶和海肾萤光素酶也经常被称作荧光素、荧光素酶、萤火虫荧光素酶和海肾荧光素酶。
      pGL6-TA主要用于在其多克隆位点插入特定的启动子、增强子等调控元件研究该调控序列的基因转录调控活性。
      pGL6-TA和pGL6相比在多克隆位点和luc基因之间加入了一段minimal TA promoter,使luc基因的基础转录水平提高。
      pGL6-TA质粒的主要信息如下:

Base pairs 5629
Multiple cloning region 1-59
Minimal TA promoter (pTA) 66-88
luc2 reporter gene 120-1782
SV40 late poly(A) signal 1817-2038
SV40 early enhancer/promoter 2086-2504
Synthetic neomycin phosphotransferase(Neor)coding region 2529-3323
Synthetic poly(A) signal 3348-3396
Reporter Vector primer 4 (RVprimer4) binding region 3463-3482
ColE1-derived plasmid replication origin 3720
Synthetic Beta-lactamase (Ampr) coding region 4511-5371
Synthetic poly(A) signal/transcriptional pause site 5476-5629
Reporter Vector primer 3 (RVprimer3) binding region 5578-5597

      pGL6-TA质粒的图谱如下:

      pGL6-TA的多克隆位点的详细图谱如下:

BglI KpnI NheI XhoI SacI MluI BglII
1 GGCCTAACTG GCCGGTACCG CTAGCCTCGA GGAGCTCACG CGTAGATCTG
CCGGATTGAC CGGCCATGGC GATCGGAGCT CCTCGAGTGC GCATCTAGAC
HindIII Minimal TA promoter
51 CAGAAGCTTA
GACAC TAGAG
GGTATATAAT
GGAAGCTC GA
CTTCCAGCTT
GTCTTCGAAT
CTGTG ATCTC
CCATATATTA
CCTTCGAG CT
GAAGGTCGAA

      pGL6-TA中没有的酶切位点(Restriction enzymes that do not cut pGL6-TA)包括:

Aat II Afl II Asc I Ase I Bsa I BsaA I BsiW I
BspM II BssH II Eco72 I EcoR I EcoR V Nde I Nru I
PflM I Pme I Pml I Psp1406 I PspA I Rsr II Sma I
SnaB I Spl I Srf I Tth111 I

      pGL6-TA中的单酶切位点(Restriction enzymes that cut pGL6-TA once)包括:

Sfi IGGCCN,NNN`NGGCC 9 Stu I AGG|CCT 2483
Bgl IGCCN,NNN`NGGC 9 EcoN I CCTNN`N,NNAGG 3004
Acc65 I G`GTAC,C 15 BsiC I TT`CG,AA 3399
Asp718 G`GTAC,C 15 BstB I TT`CG,AA 3399
Kpn I G,GTAC`C 19 Sal I G`TCGA,C 3413
Nhe I G`CTAG,C 21 ApaL I G`TGCA,C 3977
PaeR7I C`TCGA,G 27 HgiE II ACCNNNNNNGGT -1/13 4242
Xho I C`TCGA,G 27 Not I GC`GGCC,GC 4483
Sac I G,AGCT`C 37 BstX I CCAN,NNNN`NTGG 4507
Mlu I A`CGCG,T 39 BstE II G`GTNAC,C 4510
Bgl II A`GATC,T 45 Ahd I GACNN,N`NNGTC 4585
Hind III A`AGCT,T 55 Bsu36 I CC`TNA,GG 4941
BsrG I T`GTAC,A 621 Pvu I CG,AT`CG 4955
Dra III CAC,NNN`GTG 1277 Sac II CC,GC`GG 4979
Gsu I CTGGAG 21/19 1510 Bst1107 I GTA|TAC 5095
Bpm I CTGGAG 22/20 1511 Xca I GTA|TAC 5095
Apo I R`AATT,Y 1893 Spe I A`CTAG,T 5414
Mun I C`AATT,G 1957 BsmA I GTCTC`/9 5426
BamH I G`GATC,C 2050 BsmB I CGTCTC 7/11 5427

      pGL6-TA质粒中推荐使用的测序引物序列如下:
      RVprimer3(5578-5597):
      CTA GCA AAA TAG GCT GTC CC
      pGL6-TA的全序列信息:D2105-pGL6-TA-seq.txt
包装清单:

产品编号 产品名称 包装
D2105-1μg pGL6-TA (报告基因质粒) 1μg
D2105-100μg pGL6-TA (报告基因质粒) 100μg
说明书 1份

保存条件:
      -20℃保存。
注意事项:
      本质粒未经碧云天书面许可不得用于任何商业用途,也不得移交给订货人所在实验室外的任何个人或单位。
      本产品仅限于专业人员的科学研究用,不得用于临床诊断或治疗,不得用于食品或药品,不得存放于普通住宅内。
      为了您的安全和健康,请穿实验服并戴一次性手套操作。

使用说明:
1. 首次使用1μg包装的本产品时,请先取少量本质粒转化大肠杆菌,进行质粒小量、中量或大量抽提后再用于后续用途。抽提获得的质粒可以通过酶切电泳进行鉴定,或通过测序进行鉴定。
2. 100μg包装的本产品质粒浓度为0.1μg/μl,共1ml。可以直接用于酶切或者转染细胞。
3. 用于插入调控序列:在多克隆位点选取适当的酶切位点,经酶切处理后连入适当的基因转录调控序列。pGL6-TA也可以用作报告基因检测时的阴性对照。
4. pGL6-TA质粒以及以此质粒为模板构建的质粒可以用常规的细胞转染方法转染细胞。检测时可以采用碧云天的萤火虫萤光素酶报告基因检测试剂盒(RG005/RG006)或双萤光素酶报告基因检测试剂盒(RG027/RG028)。
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