| ProductName | ThermusaquaticusMutSDNAmismatchrepairprotein,N-6XHis-MBP-Taq-Muts |
| Size | 500µg |
| Description | TheTaqMutSDNAmismatchproteinrecognizesheteroduplexDNAscontainingmispairedorunpairedbases.ThisMutsproteinbindsinvitrotoheteroduplexDNAscontainingmispairedorunpairedbasesoverawidetemperaturerangefrom4to70 °CandhasaThermostableATPaseactivity.ThisthermostableTaqMutSisactiveattemperaturebetween0to75°C.SinceTaqMutSefficientlybindsto1-4basesdeletion(orinsertion)andmismatchbasepairsofGT,CTandAG,itisusefulfordetectingthesemutations.MutationscanbedetectedinpolyacrylamidegelsoronasolidphasesuchasNiagaroseoramylosebeads,ormagneticNi-NTAparticles. |
| Applications |
|
| Source | E.coli |
| FusionTag | 6XHistagandMBPtagatN-terminus |
| PurificationMethod | FPLC |
| Concentration | 1mg/ml |
| Purity | ~95%asdeterminedbySDS-PAGE
|
| Accession# | AAC43637,TAU3311,U33117 |
| GeneName | ThermusaquaticusMutSDNAmismatchrepairprotein |
| MW | 135.9kDa |
| ProteinSequence | 1MGSSHHHHHHGTKTEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKLEEKFP 61QVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTWDAVRYNGKLIAYPI 121AVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWPLIAADGGYA 181FKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAFNKGETAMTING 241PWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKELAKEFLENYLLTDE 301GLEAVNKDKPLGAVALKSYEEELAKDPRIAATMENAQKGEIMPNIPQMSAFWYAVRTAVI 361NAASGRQTVDEALKDAQTGTDYDIPTTENLYFQGHMEGMLKGEGPGPLPPLLQQYVELRD 421QYPDYLLLFQVGDFYECFGEDAERLARALGLVLTHKTSKDFTTPMAGIPLRAFEAYAERL 481LKMGFRLAVADQVEPAEEAEGLVRREVTQLLTPGTLLQESLLPREANYLAAIATGDGWGL 541AFLDVSTGEFKGTVLKSKSALYDELFRHRPAEVLLAPELLENGAFLDEFRKRFPVMLSEA 601PFEPEGEGPLALRRARGALLAYAQRTQGGALSLQPFRFYDPGAFMRLPEATLRALEVFEP 661LRGQDTLFSVLDETRTAPGRRLLQSWLRHPLLDRGPLEARLDRVEGFVREGALREGVRRL 721LYRLADLERLATRLELGRASPKDLGALRRSLQILPELRALLGEEVGLPDLSPLKEELEAA 781LVEDPPLKVSEGGLIREGYDPDLDALRAAHREGVAYFLELEERERERTGIPTLKVGYNAV 841FGYYLEVTRPYYERVPKEYRPVQTLKDRQRYTLPEMKEKEREVYRLEALIRRREEEVFLE 901VRERAKRQAEALREAARILAELDVYAALAEVAVRYGYVRPRFGDRLQIRAGRHPVVERRT 961EFVPNDLEMAHELVLITGPNMAGKSTFLRQTALIALLAQVGSFVPAEEAHLPLFDGIYTR 1021IGASDDLAGGKSTFMVEMEEVALILKEATENSLVLLDEVGRGTSSLDGVAIATAVAEALH 1081ERRAYTLFATHYFELTALGLPRLKNLHVAAREEAGGLVFYHQVLPGPASKSYGVEVAAMA 1141GLPKEVVARARALLQAMAARREGALDAVLERLLALDPDRLTPLEALRLLQELKALALGAP 1201LDTMKG |
| StorageBuffer | 20mMTris-HCl,pH8.0,250mMNaCl,0.1mMEDTA,1mMDTT,50%Glycerol |
| ReactionBuffer | 100mMKCl,50mMTris-HCl,pH8.5,5~20mMMgCl2,0.1mMEDTA,1mMDTT,2%Glycerol,65°C |
| Storage | -20to-80°C. |
| Shipping | 4°Cordryice |
Protocol (example) | 1.AfterfirstroundPCR,purifyPCRfragmentsusingQiagenQIAquickPCRpurificationkitwithelutionindH2O. 2.DilutePCRproductto250ng/µlin10mMTris–HCl,pH7.8,50mMNaClandheatto95oCfor5minfollowedbycoolingat0.1oC/sto25oC. 3.Addbindingbuffer(20mMTris–HCl,pH7.8,10mMNaCl,5mMMgCl2,1mMDTTand5%glycerol)toannealedPCRproduct,adjustDNAconcentrationto11.5ng/µl,add6XHis-MBP–MutSdimersto950nM. 4.Incubatethemixtureatroomtemperaturefor10min,thenaddanequalvolumeofamyloseresin(NEB)preequilibratedwith1Xbindingbuffer,andincubatefor30minatroomtemperature. 5.Gentlyspindownbeadsandsavesupernatantforsubsequentprocessing(secondroundPCR,cloningetc). |
| References | 1.Protein-mediatederrorcorrectionfordenovoDNAsynthesis.NucleicAcidsRes.2004Nov23;32(20):e162. 2.CorrectingerrorsinsyntheticDNAthroughconsensusshuffling.NucleicAcidsRes.2005Mar30;33(6):e55. 3.MutSasatoolformutationdetection.ActaBiochimPol.2005;52(3):575-83.Epub2005Aug4. 4.OnetubemutationdetectionusingsensitivefluorescentdyeingofMutSprotectedDNA.NucleicAcidsRes.2000Apr15;28(8):E36. 5.RapidSNPdiagnosticsusingasymmetricisothermalamplificationandanewmismatch-suppressiontechnology.NatureMethods-4,257-262(2007)doi:10.1038/nmeth1007 |
ebiomall.com
>
>
>
>
>
>
>
>
>
>
>
尽量简洁
缺点:质粒仍然较大,转染难度相对较大。具有碱基识别偏好性,局限了基因编辑的运用范围,而且会导致不同基因位点编辑效率不同。筛选仍然需要较大工作量。
http://www.nature.com/nature/journal/v520/n7546/full/nature14299.html
1、过表达目的蛋白,用以检测基因表型;
2、荧光标签标记目的蛋白,用以跟踪其细胞 定位;
3、引入野生型等位基因。
是的,确切来说是大量表达。 大肠杆菌是基因重组技术中常用的细菌,将外源目的基因(如人胰岛素基因)导入大肠杆菌后可在大肠杆菌内表达目的蛋白(如胰岛素),由于细菌繁殖速度快,通过发酵便可在短时间内获得大量胰岛素,再经多步分离、纯化便得到了药用胰岛素。
流程大概是这样的:首先获得小鼠ES细胞系,测试ES细胞嵌合入受体囊胚的能力之后根据不同基因、不同目的设计并构建打靶载体,将打靶载体转入一定数目ES细胞中,然后鉴定出带有发生正确同源重组的突变中靶ES细胞。通过显微注射或者胚胎融合的方法将经过遗传修饰的ES细胞引入受体胚胎内。经过遗传修饰的ES细胞可以发育为嵌合体动物的生殖细胞,是的经过修饰的遗传信息经生殖系遗传,从而得到带有修饰基因的突变小鼠,而后可以对其进行表型分析。

![]()
暂无品牌分类

