pFastBacHT A昆虫表达载体说明

pFastBacHT A

编号 载体名称

北京华越洋生物KCVECT130 pFastBacHT A

pFastBacHTA载体基本信息

载体名称: pFastBacHTA, pFastBac HTA, pFastBacHT A, pFastBac HT A

质粒类型: 昆虫细胞表达载体；杆状病毒表达载体

启动子: Polyhedrin

表达水平: 高

克隆方法: 多克隆位点，限制性内切酶

载体大小: 4856 bp

5' 测序引物: pFastbac-F: TATTCCGGATTATTCATACC

3' 测序引物: pFastBac-R: ACAAATGTGGTATGGCTGA

载体标签: N-His tag

载体抗性: Ampicillin 氨苄

筛选标记: Gentamicin 庆大霉素

备注: 含TEV切割位点

稳定性: 稳表达Stable

组成型: 组成型Constitutive

病毒/非病毒: 杆状病毒

pFastBacHTA载体质粒图谱和多克隆位点信息

pFastBacHTA载体简介

Bac-­‐to-­‐BacHT V ector K it 设计用于在大肠杆菌 (E. c oli) 中生成杆状病毒载体后，在 Sf9、Sf21 或 High F ive 细胞中表达和纯化带组氨酸标签的重组蛋白。 pFastBacHT 载体具

有以下特点：

⇒用于表达蛋白质的多角体强启动子

⇒用于简化克隆的三个读码框

⇒用于轻松纯化重组融合蛋白的 N-­‐端 6xHis 标签

⇒用于在蛋白质纯化后去除组氨酸标签的 TEV 蛋白酶切割位点

引用及参考文献

DEF6, a Novel Substrate for the Tec Kinase ITK, Contains a Glutamine-­‐rich Aggregation-­‐prone Region and Forms Cytoplasmic Granules that Co-­‐localize with P-­‐bodies.

Hey F, C zyzewicz N, J ones P, S ablitzky F,

J B iol C hem (2012) 287:31073-­‐31084

Product u sage: n a

Cytoskeletal c hanges r egulated b y t he P AK4 s erine/threonine k inase a re m ediated b y L IM kinase 1 a nd c ofilin.

Dan C; K elly A; B ernard O; M inden A;

J B iol C hem (2001) 276:32115-­‐32136

Product usage: pFASTBAC/HT and pFASTBAC-­‐1 vectors were used to transfect with Sf9 cells w ith C ellfectin.

Sp3 Represses Gene Expression via the Titration of Promoter-­‐specific Transcription Factors.

Kennett S arah B; M oorefield K S cott; H orowitz J onathan M;

J B iol C hem (2002) 277:9780-­‐9789

Product u sage:

Baculovirus stocks encoding Sp1, Sp3, M1, and M2 were prepared using the PCR, appropriate primers, and pCMV4-­‐Sp1/flu, pBSK-­‐Sp3/flu, pCR-­‐M1/flu, and pCR-­‐M2/flu as substrates. PCR reactions employed pGEX-­‐M2 using Taq polymerase. Amplified cDNAs were s ubcloned i n p FASTBacHTA.

pFastBacHTA载体序列

ORIGIN

1 GACGCGCCCT GTAGCGGCGC ATTAAGCGCG GCGGGTGTGG TGGTTACGCG CAGCGTGACC

61 GCTACACTTG CCAGCGCCCT AGCGCCCGCT CCTTTCGCTT TCTTCCCTTC CTTTCTCGCC 121 ACGTTCGCCG GCTTTCCCCG TCAAGCTCTA AATCGGGGGC TCCCTTTAGG GTTCCGATTT 181 AGTGCTTTAC GGCACCTCGA CCCCAAAAAA CTTGATTAGG GTGATGGTTC ACGTAGTGGG 241 CCATCGCCCT GATAGACGGT TTTTCGCCCT TTGACGTTGG AGTCCACGTT CTTTAATAGT 301 GGACTCTTGT TCCAAACTGG AACAACACTC AACCCTATCT CGGTCTATTC TTTTGATTTA 361 TAAGGGATTT TGCCGATTTC GGCCTATTGG TTAAAAAATG AGCTGATTTA ACAAAAATTT 421 AACGCGAATT TTAACAAAAT ATTAACGTTT ACAATTTCAG GTGGCACTTT TCGGGGAAAT 481 GTGCGCGGAA CCCCTATTTG TTTATTTTTC TAAATACATT CAAATATGTA TCCGCTCATG 541 AGACAATAAC CCTGATAAAT GCTTCAATAA TATTGAAAAA GGAAGAGTAT GAGTATTCAA 601 CATTTCCGTG TCGCCCTTAT TCCCTTTTTT GCGGCATTTT GCCTTCCTGT TTTTGCTCAC 661 CCAGAAACGC TGGTGAAAGT AAAAGATGCT GAAGATCAGT TGGGTGCACG AGTGGGTTAC 721 ATCGAACTGG ATCTCAACAG CGGTAAGATC CTTGAGAGTT TTCGCCCCGA AGAACGTTTT 781 CCAATGATGA GCACTTTTAA AGTTCTGCTA TGTGGCGCGG TATTATCCCG TATTGACGCC 841 GGGCAAGAGC AACTCGGTCG CCGCATACAC TATTCTCAGA ATGACTTGGT TGAGTACTCA 901 CCAGTCACAG AAAAGCATCT TACGGATGGC ATGACAGTAA GAGAATTATG CAGTGCTGCC 961 ATAACCATGA GTGATAACAC TGCGGCCAAC TTACTTCTGA CAACGATCGG AGGACCGAAG 1021 GAGCTAACCG CTTTTTTGCA CAACATGGGG GATCATGTAA CTCGCCTTGA TCGTTGGGAA 1081 CCGGAGCTGA ATGAAGCCAT ACCAAACGAC GAGCGTGACA CCACGATGCC TGTAGCAATG 1141 GCAACAACGT TGCGCAAACT ATTAACTGGC GAACTACTTA CTCTAGCTTC CCGGCAACAA 1201 TTAATAGACT GGATGGAGGC GGATAAAGTT GCAGGACCAC TTCTGCGCTC GGCCCTTCCG 1261 GCTGGCTGGT TTATTGCTGA TAAATCTGGA GCCGGTGAGC GTGGGTCTCG CGGTATCATT 1321 GCAGCACTGG GGCCAGATGG TAAGCCCTCC CGTATCGTAG TTATCTACAC GACGGGGAGT 1381 CAGGCAACTA TGGATGAACG AAATAGACAG ATCGCTGAGA TAGGTGCCTC ACTGATTAAG 1441 CATTGGTAAC TGTCAGACCA AGTTTACTCA TATATACTTT AGATTGATTT AAAACTTCAT 1501 TTTTAATTTA AAAGGATCTA GGTGAAGATC CTTTTTGATA ATCTCATGAC CAAAATCCCT 1561 TAACGTGAGT TTTCGTTCCA CTGAGCGTCA GACCCCGTAG AAAAGATCAA AGGATCTTCT 1621 TGAGATCCTT TTTTTCTGCG CGTAATCTGC TGCTTGCAAA CAAAAAAACC ACCGCTACCA 1681 GCGGTGGTTT GTTTGCCGGA TCAAGAGCTA CCAACTCTTT TTCCGAAGGT AACTGGCTTC 1741 AGCAGAGCGC AGATACCAAA TACTGTCCTT CTAGTGTAGC CGTAGTTAGG CCACCACTTC 1801 AAGAACTCTG TAGCACCGCC TACATACCTC GCTCTGCTAA TCCTGTTACC AGTGGCTGCT 1861 GCCAGTGGCG ATAAGTCGTG TCTTACCGGG TTGGACTCAA GACGATAGTT ACCGGATAAG 1921 GCGCAGCGGT CGGGCTGAAC GGGGGGTTCG TGCACACAGC CCAGCTTGGA GCGAACGACC 1981 TACACCGAAC TGAGATACCT ACAGCGTGAG CATTGAGAAA GCGCCACGCT TCCCGAAGGG 2041 AGAAAGGCGG ACAGGTATCC GGTAAGCGGC AGGGTCGGAA CAGGAGAGCG CACGAGGGAG 2101 CTTCCAGGGG GAAACGCCTG GTATCTTTAT AGTCCTGTCG GGTTTCGCCA CCTCTGACTT 2161 GAGCGTCGAT TTTTGTGATG CTCGTCAGGG GGGCGGAGCC TATGGAAAAA CGCCAGCAAC 2221 GCGGCCTTTT TACGGTTCCT GGCCTTTTGC TGGCCTTTTG CTCACATGTT CTTTCCTGCG 2281 TTATCCCCTG ATTCTGTGGA TAACCGTATT ACCGCCTTTG AGTGAGCTGA TACCGCTCGC 2341 CGCAGCCGAA CGACCGAGCG CAGCGAGTCA GTGAGCGAGG AAGCGGAAGA GCGCCTGATG 2401 CGGTATTTTC TCCTTACGCA TCTGTGCGGT ATTTCACACC GCAGACCAGC CGCGTAACCT 2461 GGCAAAATCG GTTACGGTTG AGTAATAAAT GGATGCCCTG CGTAAGCGGG TGTGGGCGGA 2521 CAATAAAGTC TTAAACTGAA CAAAATAGAT CTAAACTATG ACAATAAAGT CTTAAACTAG 2581 ACAGAATAGT TGTAAACTGA AATCAGTCCA GTTATGCTGT GAAAAAGCAT ACTGGACTTT 2641 TGTTATGGCT AAAGCAAACT CTTCATTTTC TGAAGTGCAA ATTGCCCGTC GTATTAAAGA