Transdirect insect cell 取扱説明書
Contents
1. A2. N A RI RI RI MSDS SHIMADZU 11 BIOTECH 1 1 IA 1 Insect Cell Extract ee 2 3 RS
3. 50 100 gg g mL pTD1 Vector DNA DDBJ GenBankP EMBL Accession Number AB194742 ma pTD1 Vector pTD1 Vector pTD1 5 UTR mRNA 2 pTD1 Vector ca T7 RNA polymerase promoter 240 258 UIR Mcs 3UTR PoyAlm Polyhedrin 5 untranslated region 259 304 ITerminator Multiple cloning region 305 343 3 Untranslated region 359 731 NMe PolyA region 736
4. Sf21 Spodoptera ugiperda Malacosma neustria nucleopolyhedrovirus 5 P N 292 30000 91 1 P N 292 30011 1 50 gL 40 Imsect Cell Extract 210 gL 5 80 C 19 Reaction Buffer 630 uL 1 80 C 4 mM Methionine 50 L 1 20 05 ug uLpTD1 Vector 10 wL
5. ki RO 2004 2007 Shimadzu Corporation All rights reserved 1 SHIMADZU BIOTECH
6. CS DO 5 pTD1 Vector 6 S HIMADZ BIOTECH 50 C N 5 GCAGATTGTACTGAGAGTG 3 161 179 C 5 ACAACGCACAGAATCTAGC 3 412 394 mRNA DNA DNA zg oz 1 RNase PCR DNA
7. Rs 22 66 2 RI
8. 50 C 1 N 5 GCAGATTGTACTGAGAGTG 3 161 179 2 M13 C 5 GGAAACAGCTATGACCATG 3 845 827 3 pTD1Vector C 101 2145 oe 1 183 co 52I 806 ozl 805 co T14I 769 w I 994 813 Scal 2543 Stul 234 SHIMADZU 7 BIOTECH mRNA mRNA 37 w mRNA mRNA 4 mRNA Code No AmpliScribe T7 fFlash Transcription Kit Epicentre ASF3257 AmpliScribe 5 T7 Transcription Kit Epicentre AS3107 Rn CUGA 7 vitro i Transcription Kit MEGA script T7 Hh Erato RIE TRIO 1334
9. C 5 2 Disa 1 PCR cDNA PCR 2 PCR Fidelity Terminal transferase PCR KODplus Code No KOD 201 N 5 ATG 2 3 ATG TGA ORF ACT 3 ACT CTTAAG GG 5 AA co RI C 4 PCR PCR PCR pTD1 Vector Zco RV
10. mRNA RNase mRNA RNase inhibitor Code No 2310A Promega Code No N2611 50 L 5 units mRNA mRNA mRNA mRNA SHIMADZU 17 BIOTECH mRNA NTPs mRNA mRNA
11. 0 5 g L Control DNA B DNA 7 0 5 g L Control DNA mRNA LS 7 8 mRNA 2 8 mRNA S 12 BIOTECH 14 MS 8 8 Promega 8 Galactosidase Enzyme Assay System with Reporter Lysis Buffer Code No E2000 Biospec mini 8 7 2 Promega
12. 120 20 1 Amersham Biosciences NICK Columns Code No 17 0855 01 2 NICK Columns mRNA Amersham Biosciences 3 mL 3 mL oO mRNA 100 uxL o gt Cm CS DS 400 L SHIMADZU 9 BIOTECH mRNA mRNA
13. 5UTR 3 DNA mn PCR Hoffman La Roche PCR SHIMADZU 5 BIOTECH In vitro DNA gco RV 7 ORF PCR 2 N C 5
14. Lot No 1 Lot No Insect Cell Extract 8 80 25 37 C mRNA T7 ind II or 1 TE 3M pH5 5 TE X X ol h 70 2 SHIMADZU BIOTECH 1 2
15. RI 18 MO BIOTECH 2004 42 639 641 2005 83 390 391 Ezure T et al Cell free protein synthesis system prepared from insect cells by freeze thawing Biotechnol Prog 2006 22 1570 1577 Suzuki T et al Performance of expression vector pTD1 for insect cell free translation system J Biosci Bioeng 2006 102 69 71 Suzuki T et g N terminal protein modifications in an insect cel free protein synthesis system and their identification by mass spectrometry Proteomics 2006 6 4486 4495 Suzuki T et g Preparation of N acylated proteins modified with fatty acids having a specific chain length using an insect cel free protein synthesis system Biosci Biotechnol Bioc
16. TE mRNA g L Azeo x 0 04 x 10 MS BIOTECH RI 121 C 20 25 C mRNA RI S Methionine Promega FluoroTectrw Greeni in vitro Translation Labeling System Code No L5001 50 gL 5 50 L mRNA 16 g Reaction Buffer 15 L Insect Cell Extract 25 UL 4 mM Methionine 1 L 50 L SHIMADZU 11 BIOTECH
17. N Promega Code No Y4041 pTD1 2 C PCR 7 DNA mRNA SS 8 mRNA mi gr6 Transdirect seo ce 11 ES 52 SHIMADZU 3 BIOTECH In vitro DNA o DNA pTD1 Vector pTD1 Vector 2 pTD1 Vector mRNA T7 5 5 UTR MCS mRNA
18. 16 NG BIOTECH mRNA DNA DH5 g M nvitrogen Code No 18258 012 LB 70 EF 7 mRNA DNA RNase InsectceuExtract Reaction Buffer 80 4 mM Methionine 50 g L pTD1 Vector 0 5 g L Control DNA 20 C EF 1
19. RI RI SDS PAGE 1 SDS PAGE CBB 2 CBB 3 X 6 1 Amersham Biosciences Amplify Fluorographic Reagents Code No NAMP100 80 C Promega FluoroTect Green in vitro Translation Labeling System Code No L5001 SDS PAGE
20. kn 4 C 7 Insect Cell Extract 80 C 30 80 C Reaction Buffer 4 mM Methionine mRNA 25 C 5 oo wh BIOTECH 12 MS 33s Methionine Promega FluoroTect Green in vitro Translation Labeling System Code No
21. ES 8 4 8 80 C FW 2 cDNA ORF Eco RV SS 5 0 5 ug uL pTD1 Vector RI a oe 3 DI
22. 1 5 mL 8 400 uL mRNA 400 L 9 mRNA 40 L 950 uxL 15 000 rpm 4 C 20 70 70 6 mRNA 20 100 wL sn mRNA 2 g L 7 mRNA 400 uL 2 CS 9 9 10 tm NTPs mRNA mRNA mRNA TE 100 400
23. 10 gL 1 x Reporter Lysis Buffer 490 L 50 50 10 uL 1 x Reporter Lysis Buffer 140 L Assay 2 Xx Buffer 150 L 37 C 30 30 1M Sodium Carbonate 500 L 420 nm 50 U mL mp 10 U mL 8 SHIMADZU 15 BIOTECH X Gal 20 mg mL X 3 L X Gal 1 L 16 L Total 20 uL 7 37 C 20 2 mRNA
24. 1 20 8 05 g L Control DNA 10 Ll 1 nr DNA Insect Cell Extract Reaction Buffer 80 C 4 mM Methionine 0 5 g LpTD1 Vector 0 5 g L Control DNA 20 80 S Methionine RI Reaction Buffer SHIMADZU 1 BIOTECH Insect Cell Extract 1 Lot No 2 Lot 3 Insect Cell Extract COz CO gt
25. L5001 50 xL Ri H Leucine 4 mM Methionine 6 50 xL 5S Met FluoroTect W mRNA 16 ug 16 ug Reaction Buffer 15 wuL 15 L Insect Cell Extract 25 L 25 uL S Methionine 1 gL 1200 Ci mmoL 10 mCi mL 4 mM Methionine 1 L FluoroTect WM Green tRNA 1 L 50 L RI RI RI S 12 1 5 SHIMADZU 13 BIOTECH
26. 757 T7 transcription terminator 758 805 ColE1 ori 835 1104 lactamase Amp coding region 1992 2852 pTD1 Vector 3052 bp 2 pTD1 Vector 4 MNO BIOTECH GCAGATTGTA CTGAGAGTGC _ ACCATATGCG GTGTGAAATA _ CCGCACAGAT GCGTAAGGAG 220 Forward primer AAAATACCGC ATCAGGCC AATACGACTC ACTATAGGIAG _ TATTTTTATT CTTTCGTAAA 280 T7 Promoter Polyhedrin PUTR Smal ES AAAATTAGAA AAATAAAATA TAAAGATATC GAATTCGAGC TCGGTACCCG GGGATCCTCT 340 i a Eco RV EcoRI Sacl Ap 1 az7 HI Xbal AGAGTCGGGC _ GGCTGTAAAA CACGATACAT TGTTATTAGT ACATTTATTA AGCGCTAGAT 400 dd TCTGTGCGTT GTTGATTTAC 420 Reverse primer 3 pTD1 Vector pTD1 Vector 5UTR 5UTR Eco RV UTR PCR
27. RiboMAX Large Scale RNA Production System T7 Promega P1300 RNAMaxx Fieh Vield Ta GG it ralagens 200339 ScriptMAX gt Thermo T7 Transcription Kit TSK101 T7 RiboMAX Express Large Scale RNA Production System Promega P1320 mRNA NTPs B mR A NTPs mRNA A A SS 9 B ES 10 8 ha BIOTECH 3M pH 5 5 2 70
28. S nD 3 4 7 vitro DNA 4 pTDl Vector en RR GSR ae RNA ENKKRKNNN KEINIIIININNOQOQNRN IE BNAC EL mm ET T ToTT me momomhmhitbooio 7 mRNA 8 mRNA 8 mRNA gg 10 HH 11 VENI 11 13 RI ee 14 14 B 14 8 i kdA Nh 15 17 17 19 MSDS Lk 19 Lk I 19 IV SHIMADZU BIOTECH Transdirect secf ce mRNA
29. SHIMAD zu OBioTEcH Transdirect insect cell 292 30011B Feb 2007 E WO Ce RR 1 2 3
30. hem 2007 71 261 264 MSDS MSDS Web 604 8511 1 TEL 075 823 1351 FAX 075 823 1364 E MAIL t direct shimadzu biotechJjp http www shimadzu biotechJp SHIMADZU 19 BIOTEC CH
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