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Fusion Proteins

When each of these genes is cloned into the pQE-30 UA vector, a fusion protein will be made beginning with six histidine amino acids and then continuing with the correct number of amino acids in the given protein. Each protein will contain the following amino acid sequences (Cherry et al., 2000):

BLUE sequences = RGS 6x Tag; GREEN sequence = polylinkers

IDH1:

Full Length Translation Product: 360 amino acids, 39326 Da

Mature Protein: 349 amino acids, 38025 Da

RGS HHHHHH GSHVISSIAS MLNRTIAKRTLATAAQAERTLPKKYGGRFTVTLIPGDGVGKEIT DSVRTIFEAENIPIDWETINIKQTDHKEGVYEAVESLKRNKIGLKGLWHTPADQTGHG SLNVALRKQLDIYANVALFKSLKGVKTRIPDIDLIVIRENTEGEFSGLEHESVPGVVE SLKVMTRPKTERIARFAFDFAKKYNRKSVTAVHKANIMKLGDGLFRNIITEIGQKEYP DIDVSSIIVDNASMQAVAKPHQFDVLVTPSMYGTILGNIGAALIGGPGLVAGANFGRD YAVFEPGSRHVGLDIKGQNVANPTAMILSSTLMLNHLGLNEYATRISKAVHETIAEGK HTTRDIGGSSSTTDFTNEIINKLSTM

IDH2:

Full Length Translation Product: 369 amino acids, 39741 Da

Mature Protein: 354 amino acids, 3796 Da

RGS HHHHHH GSHVISSIAS MLRNTFFRNTSRRFLATVKQPSIGRYTGKPNPSTGKYTVSFIEG DGIGPEISKSVKKIFSAANVPIEWESCDVSPIFVNGLTTIPDPAVQSITKNLVALKGP LATPIGKGHRSLNLTLRKTFGLFANVRPAKSIEGFKTTYENVDLVLIRENTEGEYSGI EHIVCPGVVQSIKLITRDASERVIRYAFEYARAIGRPRVIVVHKSTIQRLADGLFVNV AKELSKEYPDLTLETELIDNSVLKVVTNPSAYTDAVSVCPNLYGDILSDLNSGLSAGS LGLTPSANIGHKISIFEAVHGSAPDIAGQDKANPTALLLSSVMMLNHMGLTNHADQIQ NAVLSTIASGPENRTGDLAGTATTSSFTEAVIKRL

IDP1:

Full Length Translation Product: 428 amino acids, 48195 Da

Mature Protein: 412 amino acids, 46383 Da

RGS HHHHHH GSHVISSIAS MSMLSRRLFSTSRLAAFSKIKVKQPVVELDGDEMTRIIWDKIKK KLILPYLDVDLKYYDLSVESRDATSDKITQDAAEAIKKYGVGIKCATITPDEARVKEF NLHKMWKSPNGTIRNILGGTVFREPIVIPRIPRLVPRWEKPIIIGRHAHGDQYKATDT LIPGPGSLELVYKPSDPTTAQPQTLKVYDYKGSGVAMAMYNTDESIEGFAHSSFKLAI DKKLNLFLSTKNTILKKYDGRFKDIFQEVYEAQYKSKFEQLGIHYEHRLIDDMVAQMI KSKGGFIMALKNYDGDVQSDIVAQGFGSLGLMTSILVTPDGKTFESEAAHGTVTRHYR KYQKGEETSTNSIASIFAWSRGLLKRGELDNTPALCKFANILESATLNTVQQDGIMTK DLALACGNNERSAYVTTEEFLDAVEKRLQKEIKSIE

IDP2:

Full Length Translation Product: 412 amino acids, 46553 Da

Mature Protein: 411 amino acids, 46422 Da

RGS HHHHHH GSHVISSIAS MTKIKVANPIVEMDGDEQTRIIWHLIRDKLVLPYLDVDLKYYDL SVEYRDQTNDQVTVDSATATLKYGVAVKCATITPDEARVEEFHLKKMWKSPNGTIRNI LGGTVFREPIIIPRIPRLVPQWEKPIIIGRHAFGDQYKATDVIVPEEGELRLVYKSKS GTHDVDLKVFDYPEHGGVAMMMYNTTDSIEGFAKASFELAIERKLPLYSTTKNTILKK YDGKFKDVFEAMYARSYKEKFESLGIWYEHRLIDDMVAQMLKSKGGYIIAMKNYDGDV ESDIVAQGFGSLGLMTSVLITPDGKTFESEAAHGTVTRHFRQHQQGKETSTNSIASIF AWTRGIIQRGKLDNTPDVVKFGQILESATVNTVQEDGIMTKDLALILGKSERSAYVTT EEFIDAVESRLKKEFEAAAL

IDP3:

Full Length Translation Product: 420 amino acids, 47866 Da

Mature Protein: 419 amino acids, 47734 Da

RGS HHHHHH GSHVISSIAS MSKIKVVHPIVEMDGDEQTRVIWKLIKEKLILPYLDVDLKYYDL SIQERDRTNDQVTKDSSYATLKYGVAVKCATITPDEARMKEFNLKEMWKSPNGTIRNI LGGTVFREPIIIPKIPRLVPHWEKPIIIGRHAFGDQYRATDIKIKKAGKLRLQFSSDD GKENIDLKVYEFPKSGGIAMAMFNTNDSIKGFAKASFELALKRKLPLFFTTKNTILKN YDNQFKQIFDNLFDKEYKEKFQALKITYEHRLIDDMVAQMLKSKGGFIIAMKNYDGDV QSDIVAQGFGSLGLMTSILITPDGKTFESEAAHGTVTRHFRKHQRGEETSTNSIASIF AWTRAIIQRGKLDNTDDVIKFGNLLEKATLDTVQVGGKMTKDLALMLGKTNRSSYVTT EEFIDEVAKRLQNMMLSSNEDKKGMCKL

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