Protein yog ib qho tseem ceeb ntawm txhua yam kab mob. Txhua yam ntawm nws cov molecules muaj ib los yog ntau dua polypeptide chains uas muaj cov amino acids. Txawm hais tias cov ntaub ntawv tsim nyog rau lub neej yog encoded nyob rau hauv DNA los yog RNA, recombinant proteins ua ib tug ntau yam ntawm lom functions nyob rau hauv cov kab mob, nrog rau enzymatic catalysis, kev tiv thaiv, kev txhawb nqa, txav, thiab kev tswj. Raws li lawv cov dej num hauv lub cev, cov tshuaj no tuaj yeem muab faib ua ntau pawg, xws li cov tshuaj tiv thaiv kab mob, cov enzymes, cov khoom tiv thaiv. Muab lawv lub luag haujlwm tseem ceeb, cov tebchaw no tau raug tshawb fawb thiab siv dav.
Yav dhau los, txoj hauv kev tseem ceeb kom tau txais cov protein sib xyaw ua ke yog cais tawm ntawm lub ntuj tsim, uas feem ntau tsis muaj txiaj ntsig thiab siv sijhawm. Kev nce qib tsis ntev los no hauv cov thev naus laus zis molecular tau ua rau nws muaj peev xwm los clone DNA encoding ib txheej tshwj xeeb ntawm cov tshuaj mus rau hauv ib qho kev qhia vector rau cov khoom xws li cov kab mob, cov poov xab, kab kab, thiab cov kab mob mammalian.
Y Yuav muab tso yooj yim, cov proteins recombinant tau txhais los ntawm exogenous DNA khoom rau hauvnyob hlwb. Tau txais lawv feem ntau suav nrog ob kauj ruam tseem ceeb:
- Cloning ib molecule.
- YProtein expression.
Tam sim no, kev tsim cov qauv zoo li no yog ib txoj hauv kev muaj zog tshaj plaws siv hauv tshuaj thiab biology. Cov khoom xyaw muaj ntau daim ntawv thov hauv kev tshawb fawb thiab biotechnology.
kev taw qhia kho mob
Cov proteins sib xyaw ua ke muab kev kho mob tseem ceeb rau ntau yam kab mob xws li ntshav qab zib, mob qog noj ntshav, kab mob sib kis, hemophilia thiab ntshav ntshav. Cov qauv tsim ntawm cov tshuaj xws li tshuaj tiv thaiv kab mob, tshuaj hormones, interleukins, enzymes, thiab anticoagulants. Muaj kev xav tau ntau ntxiv rau cov tshuaj sib xyaw ua ke rau kev siv kho mob. Lawv tso cai rau koj nthuav txoj kev kho mob.
cov noob caj noob ces rov ua cov proteins ua lub luag haujlwm tseem ceeb hauv kev lag luam tshuaj kho mob. Mammalian hlwb tam sim no tsim cov tshuaj kho mob tshaj plaws vim lawv cov formulations muaj peev xwm ua tau zoo, zoo li tej yam ntuj tso. Tsis tas li ntawd, ntau qhov kev pom zoo kho cov proteins uas tau tsim nyob rau hauv E. coli vim yog cov caj ces zoo, kev loj hlob sai, thiab kev tsim khoom siab. Nws kuj muaj txiaj ntsig zoo rau kev tsim cov tshuaj raws li cov tshuaj no.
Kev Tshawb Fawb
Tau txais cov protein ntau dua yog nyob ntawm ntau txoj kev. Cov tshuaj pab kom paub cov ntsiab lus tseem ceeb ntawm lub cev. Cov molecules no tuaj yeem siv los txheeb xyuas thiab txiav txim siabqhov chaw ntawm cov tshuaj encoded los ntawm ib tug tshwj xeeb gene, thiab qhia txog kev ua hauj lwm ntawm lwm cov noob nyob rau hauv ntau yam cellular kev ua ub no xws li xov tooj ntawm tes signaling, metabolism, kev loj hlob, replication thiab tuag, transcription, txhais thiab hloov kho cov tebchaw uas tau tham nyob rau hauv tsab xov xwm.
Yog li, qhov pom muaj nyob hauv feem ntau siv rau hauv molecular biology, cell biology, biochemistry, structural thiab biophysical kev tshawb fawb thiab ntau lwm yam kev tshawb fawb. Nyob rau tib lub sijhawm, tau txais cov protein sib xyaw ua ke yog kev xyaum thoob ntiaj teb.
Cov tebchaw no yog cov cuab yeej tseem ceeb hauv kev nkag siab txog kev sib cuam tshuam ntawm tes. Lawv tau ua pov thawj zoo hauv ntau txoj hauv kev kuaj xws li ELISA thiab immunohistochemistry (IHC). Recombinant proteins tuaj yeem siv los tsim cov tshuaj enzyme. Thaum siv ua ke nrog ib khub ntawm cov tshuaj tiv thaiv tsim nyog, cov hlwb tuaj yeem siv los ua cov qauv rau cov thev naus laus zis tshiab.
YBiotechnology
Recombinant proteins uas muaj cov amino acid ib ntus kuj tau siv hauv kev lag luam, khoom noj khoom haus, kev ua liaj ua teb thiab bioengineering. Piv txwv li, hauv kev yug tsiaj, enzymes tuaj yeem muab ntxiv rau hauv cov zaub mov kom muaj txiaj ntsig zoo ntawm cov khoom noj pub, txo cov nqi thiab pov tseg, pab tsiaj plab noj qab haus huv, txhim kho kev tsim khoom, thiab txhim kho ib puag ncig.
Ntxiv rau, cov kab mob lactic acid (LAB) tau ntevtau siv los tsim cov khoom noj fermented, thiab tsis ntev los no LAB tau tsim los rau kev qhia txog cov proteins uas muaj cov amino acid ib ntus, uas tuaj yeem siv dav, piv txwv li, txhawm rau txhim kho tib neeg, tsiaj txhu thiab kev zom zaub mov.
Txawm li cas los xij, cov tshuaj no kuj muaj kev txwv:
- Qee zaum, kev tsim cov proteins sib xyaw ua ke yog qhov nyuaj, raug nqi thiab siv sijhawm.
- Cov khoom tsim tawm hauv cov hlwb yuav tsis sib xws cov ntaub ntawv ntuj. Qhov sib txawv no tuaj yeem txo cov txiaj ntsig ntawm kev kho cov proteins recombinant thiab txawm ua rau muaj kev phiv. Tsis tas li ntawd, qhov sib txawv no tuaj yeem cuam tshuam cov txiaj ntsig ntawm kev sim.
- Qhov teeb meem tseem ceeb ntawm txhua cov tshuaj recombinant yog immunogenicity. Txhua yam khoom siv biotech tuaj yeem pom qee yam ntawm kev tiv thaiv kab mob. Nws yog ib qho nyuaj rau kwv yees qhov kev nyab xeeb ntawm cov protein kho tshiab.
Feem ntau, kev nce qib hauv biotechnology tau nce thiab pab txhawb kev tsim cov proteins sib xyaw rau ntau yam kev siv. Txawm hais tias lawv tseem muaj qee qhov tsis zoo, cov tshuaj tseem ceeb hauv tshuaj, kev tshawb fawb thiab biotechnology.
kab mob sib kis
recombinant protein tsis muaj teeb meem rau tib neeg. Nws tsuas yog ib feem tseem ceeb ntawm tag nrho cov molecule hauv kev txhim kho cov tshuaj tshwj xeeb lossis cov khoom noj khoom haus. Ntau qhov kev tshawb fawb kho mob tau pom tias kev yuam kev ntawm FGFBP3 protein (abbreviated BP3) nyob rau hauv ib qho chaw kuaj mob ntawm cov nas rog rog pom qhov txo qis hauv lawv lub cev rog.huab hwm coj, txawm hais tias cov noob caj noob ces siv.
Cov txiaj ntsig ntawm cov kev sim no qhia tau hais tias FGFBP3 protein tuaj yeem muab kev kho tshiab rau cov kab mob metabolic xws li mob ntshav qab zib hom 2 thiab kab mob siab rog. Tab sis vim hais tias BP3 yog ib qho protein ntau thiab tsis yog cov tshuaj siv dag zog, kev sim tshuaj ntsuam xyuas ntawm tib neeg cov BP3 tuaj yeem pib tom qab qhov kawg ntawm kev tshawb fawb preclinical. Ntawm, uas yog, muaj cov laj thawj ntsig txog kev nyab xeeb ntawm kev ua cov kev tshawb fawb no. Cov protein recombinant tsis muaj teeb meem rau tib neeg vim nws cov txheej txheem ua kom huv thiab huv. Kev hloov pauv tau tshwm sim ntawm qib molecular ib yam nkaus.
PD-L2, yog ib tus neeg tseem ceeb hauv kev tiv thaiv kab mob, tau raug xaiv tsa rau 2018 Nobel nqi zog hauv Physiology lossis Tshuaj. Txoj haujlwm no, pib los ntawm Prof. James P. Allison los ntawm Tebchaws Meskas thiab Prof. Tasuku Honjo los ntawm Nyij Pooj, tau coj mus rau kev kho mob qog noj ntshav xws li melanoma, mob ntsws cancer, thiab lwm yam raws li kev kuaj xyuas cov tshuaj tiv thaiv kab mob. Tsis ntev los no, AMSBIO tau ntxiv cov khoom tshiab loj rau nws cov kab mob tiv thaiv kab mob, PD-L2 / TCR activator - CHO Recombinant Cell Line.
Nyob rau hauv cov ntaub ntawv pov thawj-ntawm-lub tswv yim kev sim, cov kws tshawb fawb ntawm University of Alabama ntawm Birmingham, coj los ntawm H. Long Zheng, MD, xibfwb Robert B. Adams, thiab Tus Thawj Coj ntawm Laboratory Medicine, Department of Pathology, UAB Tsev Kawm Ntawv ntawm Cov tshuaj, tau hais txog qhov kev kho mob uas tsis tshua muaj tab sis tuag los ntshav, TTP.
Qhov tshwm sim ntawm qhov noCov kev tshawb fawb pom thawj zaug tias kev hloov pauv ntawm rADAMTS13-loaded platelets tuaj yeem yog qhov tshiab thiab muaj peev xwm kho tau zoo rau cov hlab ntsha ntawm cov hlab ntsha cuam tshuam nrog kev yug me nyuam thiab kev tiv thaiv kab mob TTP.
Recombinant protein tsis yog tsuas yog ib qho khoom noj xwb, tab sis kuj yog ib qho tshuaj hauv cov tshuaj uas tsim los. Cov no tsuas yog ob peb thaj chaw uas tam sim no koom nrog hauv tshuaj thiab cuam tshuam nrog kev kawm ntawm tag nrho nws cov qauv tsim. Raws li kev xyaum qhia thoob ntiaj teb, cov qauv ntawm cov khoom ua rau nws muaj peev xwm ntawm qib molecular los daws ntau yam teeb meem loj hauv tib neeg lub cev.
Kev tsim tshuaj tiv thaiv
A recombinant protein yog ib qho tshwj xeeb ntawm cov molecules uas tuaj yeem ua qauv. Cov cuab yeej zoo sib xws yog siv los tsim cov tshuaj tiv thaiv. Lub tswv yim tshuaj tiv thaiv tshiab, tseem hu ua kev siv tshuaj tiv thaiv kab mob tshwj xeeb, tuaj yeem tiv thaiv ntau lab tus qaib uas muaj kev pheej hmoo los ntawm kev mob ua pa hnyav, cov kws tshawb fawb los ntawm University of Edinburgh thiab lub koom haum Pirbright tau hais. Cov tshuaj tiv thaiv no siv tsis muaj teeb meem lossis tsis muaj zog ntawm tus kab mob lossis kab mob los qhia cov kab mob rau hauv lub cev lub hlwb. Hauv qhov no, cov kws tshaj lij tau siv cov kab mob sib kis nrog cov kab mob sib txawv sib txawv ua cov tshuaj tiv thaiv los tsim ob hom kab mob uas tsis muaj mob. Muaj ntau yam tshuaj sib txawv tsim nyob ib ncig ntawm qhov kev sib txuas no.
Recombinant protein luam npe thiab cov analogues yog raws li hauv qab no:
- "Fortelizin".
- "Z altrap".
- "Eylea".
Cov no yog cov tshuaj tiv thaiv kabmob kheesxaws, tab sis muaj lwm yam kev kho mob cuam tshuam nrog cov tshuaj no.
Ib tshuaj tiv thaiv tshiab, tseem hu ua LASSARAB, tsim los tiv thaiv tib neeg tawm tsam Lassa fever thiab rabies, tau pom cov txiaj ntsig tau zoo hauv kev tshawb fawb ua ntej, raws li kev tshawb fawb tshiab luam tawm hauv phau ntawv xov xwm tshawb fawb Nature Communications. Tus neeg sib tw txhaj tshuaj tiv thaiv tsis tau rov ua dua siv tus kab mob rabies tsis muaj zog.
Pab pawg tshawb fawb tau tso cov kab mob Lassa genetic khoom mus rau hauv tus kab mob rabies vector kom cov tshuaj tiv thaiv yuav nthuav tawm cov protein ntau hauv ob lub Lassa thiab rabies hlwb. Cov khoom sib xyaw saum npoo no ua rau lub cev tiv thaiv kab mob tiv thaiv kab mob. Cov tshuaj tiv thaiv no tau siv tsis tau los "ua kom puas" tus kab mob rabies uas muaj sia nyob siv los ua tus cab.
Txoj Kev Taug Kev
Muaj ntau lub tshuab tsim khoom. Txoj kev dav dav kom tau txais cov protein sib xyaw ua ke yog raws li tau txais cov khoom siv lom neeg los ntawm kev sib txuas. Tab sis muaj lwm txoj hauv kev.
Tam sim no muaj tsib lub ntsiab lus qhia:
- E. coli expression system.
- Yeast expression system.
- Kab cell qhia qhov system.
- Mammalian cell expression system.
- Cell-free protein expression system.
Qhov kev xaiv tom kawg yog qhov tshwj xeeb tshaj yog tsim nyog rau kev nthuav qhia ntawm cov proteins transmembranethiab toxic compounds. Nyob rau hauv xyoo tas los no, cov khoom uas nyuaj rau kev nthuav qhia los ntawm cov txheej txheem intracellular tau ua tiav rau hauv cov hlwb hauv vitro. Nyob rau hauv Belarus, zus tau tej cov recombinant proteins yog dav siv. Muaj ntau lub xeev cov tuam txhab lag luam cuam tshuam txog qhov teeb meem no.
Cell Free Protein Synthesis System yog txoj hauv kev nrawm thiab muaj txiaj ntsig rau kev sib txuas cov hom phiaj los ntawm kev ntxiv ntau yam substrates thiab lub zog sib txuas tsim nyog rau kev sau thiab txhais lus hauv cov txheej txheem enzymatic ntawm cov cellular rho tawm. Nyob rau hauv xyoo tas los no, qhov zoo ntawm cov txheej txheem cell-free rau hom tshuaj xws li complex, toxic membranes tau maj mam tshwm sim, qhia lawv muaj peev xwm siv nyob rau hauv lub biopharmaceutical teb.
Cell-free thev naus laus zis tuaj yeem ntxiv ntau yam uas tsis yog qhov tshwm sim ntawm cov amino acids tau yooj yim thiab tswj tau kom ua tiav cov txheej txheem hloov kho nyuaj uas nyuaj rau kev daws teeb meem tom qab cov lus pom zoo sib xws. Cov txheej txheem zoo li no muaj txiaj ntsig zoo rau kev xa tshuaj thiab kev tsim tshuaj tiv thaiv uas siv cov kab mob zoo li cov kab mob. Ib tug loj tus naj npawb ntawm membrane proteins tau ntse qhia nyob rau hauv dawb hlwb.
Qhia tawm kev sau
Recombinant protein CFP10-ESAT 6 yog tsim thiab siv los tsim cov tshuaj tiv thaiv. Xws li tuberculosis allergen tso cai rau koj kom muaj zog tiv thaiv kab mob thiab tsim cov tshuaj tiv thaiv. Feem ntau, cov kev tshawb fawb molecular koom nrog kev kawm txog txhua yam ntawm cov protein, xws li cov qauv, kev ua haujlwm, kev hloov kho, kev ua haujlwm hauv cheeb tsam, lossis kev sib cuam tshuam. Tshawb nrhiavYuav ua li cas cov tshuaj tshwj xeeb tswj cov txheej txheem sab hauv, cov kws tshawb fawb feem ntau xav tau txoj hauv kev los tsim cov khoom sib xyaw ua haujlwm ntawm kev txaus siab thiab txiaj ntsig.
Muab qhov loj thiab qhov nyuaj ntawm cov proteins, tshuaj synthesis tsis yog ib qho kev xaiv siv tau rau qhov kev sim no. Hloov chaw, cov hlwb nyob thiab lawv cov tshuab cellular feem ntau yog siv los ua cov chaw tsim khoom thiab tsim cov tshuaj raws li cov qauv ntawm cov noob caj noob ces. Lub recombinant protein qhia system ces tsim cov qauv tsim nyog los tsim cov tshuaj. Tom ntej no los ntawm kev xaiv cov khoom tsim nyog rau ntau hom tshuaj.
Tsis zoo li cov proteins, DNA yog ib qho yooj yim los tsim cov khoom siv hluavtaws lossis hauv vitro siv cov txheej txheem tsim kom zoo. Yog li ntawd, DNA qauv ntawm cov noob caj noob ces, nrog lossis tsis ntxiv cov neeg sau xov xwm kab ke lossis kev sib raug zoo tag ib ntus, tuaj yeem tsim los ua cov qauv rau kev qhia txog cov tshuaj saib xyuas. Xws li cov tebchaw muab los ntawm cov qauv DNA no hu ua cov proteins sib xyaw ua ke.
Cov tswv yim ib txwm siv rau kev qhia txog cov khoom muaj feem cuam tshuam cov hlwb nrog DNA vector uas muaj cov qauv thiab tom qab ntawd ua rau cov hlwb los sau thiab txhais cov protein uas xav tau. Feem ntau, cov hlwb tom qab ntawd lysed kom rho tawm cov lus qhia rau kev ua kom huv tom qab. Cov recombinant protein CFP10-ESAT6 yog ua tiav nyob rau hauv txoj kev no thiab mus los ntawm ib tug purification system los ntawm ua tautsim cov co toxins. Tsuas yog tom qab ntawd nws nkag mus rau hauv cov tshuaj tiv thaiv.
Ob leeg prokaryotic thiab eukaryotic hauv vivo qhia tshuab rau cov tshuaj molecular tau siv dav. Qhov kev xaiv ntawm qhov system nyob ntawm hom protein, qhov yuav tsum tau ua rau kev ua haujlwm, thiab qhov xav tau tawm los. Cov kab lus no muaj xws li tsiaj txhu, kab, kab mob, kab mob, algae thiab hlwb. Txhua qhov system muaj nws tus kheej zoo thiab cov nyom, thiab xaiv txoj cai rau ib daim ntawv thov tshwj xeeb yog qhov tseem ceeb rau kev ua tiav ntawm cov khoom nyob rau hauv kev tshuaj xyuas.
Qhia tawm ntawm cov tsiaj nyeg
Kev siv cov proteins sib xyaw ua ke tso cai rau kev tsim cov tshuaj tiv thaiv thiab tshuaj ntawm ntau qib. Rau qhov no, txoj kev tau txais cov khoom siv no tuaj yeem siv. Mammalian kev qhia tshuab tuaj yeem siv los tsim cov proteins los ntawm tsiaj lub nceeg vaj uas muaj cov qauv thiab kev ua haujlwm ntau tshaj plaws vim lawv lub cev muaj feem cuam tshuam ib puag ncig. Qhov no ua rau muaj kev ua haujlwm siab tom qab kev txhais lus thiab kev ua haujlwm. Mammalian kev qhia tshuab tuaj yeem siv los tsim cov tshuaj tiv thaiv kab mob, cov protein ntau, thiab cov tebchaw rau kev siv hauv kev soj ntsuam ntawm tes. Txawm li cas los xij, cov txiaj ntsig no tau txuas nrog cov kab lis kev cai nruj dua.
Mammalian kev qhia tshuab tuaj yeem siv los tsim cov proteins ib ntus lossis los ntawm cov kab ntawm tes ruaj khov uas qhov kev nthuav qhia tsim tau koom ua ke rau hauv tus tswv tsev genome. Thaum cov tshuab zoo li no tuaj yeem siv rau hauv ntau qhov kev sim, lub sijhawmkev tsim khoom tuaj yeem tsim kom muaj ntau yam khoom hauv ib mus rau ob lub lis piam. Hom recombinant protein biotechnology no yog nyob rau hauv siab thov.
Cov kev hloov pauv no, cov txiaj ntsig zoo ntawm cov tsiaj nyeg siv cov kab lis kev cai raug tshem tawm thiab tuaj yeem tawm tau grams ib liter. Tsis tas li ntawd, cov proteins no muaj ntau haiv neeg folding thiab tom qab kev hloov pauv hloov pauv xws li glycosylation piv rau lwm cov kab lus.
Kab qhia
Txoj kev tsim cov protein sib xyaw tsis txwv rau cov tsiaj txhu. Kuj tseem muaj ntau txoj hauv kev tsim cov nqi tsim khoom, txawm hais tias cov txiaj ntsig ntawm cov tshuaj ib 1 liter ntawm cov kua kho tau qis dua.
Kab kab tuaj yeem siv los qhia txog qib protein ntau nrog kev hloov kho zoo ib yam li cov kab mob mammalian. Muaj ntau lub tshuab uas tuaj yeem siv los tsim cov kab mob baculovirus rov qab, uas tuaj yeem siv los rho tawm cov khoom txaus siab rau cov kab kab.
Kev nthuav qhia ntawm cov proteins sib xyaw ua ke tuaj yeem yooj yim scaled thiab yoog rau cov kab lis kev cai siab ceev ncua kev kawm rau kev sib xyaw ua ke ntawm cov molecules loj. Lawv yog cov functionality ntau zoo ib yam li haiv neeg muaj pes tsawg leeg ntawm mammalian teeb meem. Txawm hais tias cov qoob loo tuaj yeem nce txog 500 mg / L, kev tsim cov kab mob baculovirus tuaj yeem siv sijhawm ntev thiab kab lis kev cai nyuaj dua li cov kab mob prokaryotic. Txawm li cas los xij, hauv ntau lub teb chaws yav qab teb thiab sov, zoo ib yamtxoj kev suav hais tias ua tau zoo dua.
Bacterial expression
Kev tsim cov protein ntau tuaj yeem tsim nrog kev pab ntawm cov kab mob. Cov cuab yeej no txawv ntau ntawm cov uas tau piav saum toj no. Bacterial protein qhia tshuab yog nrov vim hais tias cov kab mob no yooj yim rau kab lis kev cai, loj hlob sai, thiab muab siab yields ntawm recombinant formulation. Txawm li cas los xij, ntau cov tshuaj eukaryotic uas qhia hauv cov kab mob feem ntau tsis ua haujlwm vim tias cov hlwb tsis tau nruab los ua qhov tsim nyog tom qab kev hloov pauv hloov lossis molecular folding.
Ntxiv rau, ntau cov proteins dhau los ua cov molecules suav nrog, uas nyuaj heev kom rov zoo yam tsis muaj qhov hnyav denaturators thiab cov txheej txheem tom qab cumbersome molecular refolding. Txoj kev no feem ntau suav hais tias yog tseem muaj kev sim ntau.
Cell free expression
Recombinant protein uas muaj cov amino acid ib ntus ntawm staphylokinase tau txais los ntawm txoj kev sib txawv me ntsis. Nws suav nrog ntau hom kev txhaj tshuaj, xav tau ntau lub tshuab ua ntej siv.
Cell free protein qhia yog ib qho hauv vitro synthesis ntawm cov khoom siv translationally tshaj tag nrho cov cell rho tawm. Raws li txoj cai, tag nrho cov cell rho tawm muaj tag nrho cov macromolecules thiab cov khoom uas xav tau rau kev sau, txhais lus, thiab txawm tias tom qab kev hloov pauv.
Cov khoom no suav nrog RNA polymerase, tswj hwm cov protein ntau, cov ntaub ntawv hloov pauv, ribosomes thiab tRNA. Thaum ntxivcofactors, nucleotides thiab ib hom kab mob tshwj xeeb, cov tshuaj rho tawm tuaj yeem tsim cov proteins uas txaus siab rau ob peb teev.
Txawm tias tsis muaj kev ruaj ntseg rau kev tsim khoom loj, cell-dawb lossis hauv vitro protein qhia (IVT) cov tshuab muaj ntau qhov zoo dua li cov qauv hauv vivo systems.
Cell-free qhia tso cai sai synthesis ntawm recombinant formulations tsis muaj cell kab lis kev cai. Cell-dawb systems ua rau nws muaj peev xwm sau cov proteins nrog hloov pauv cov amino acids, nrog rau kev nthuav qhia cov tebchaw uas ua rau muaj kev puas tsuaj sai sai los ntawm intracellular proteases. Tsis tas li ntawd, nws yooj yim dua los nthuav qhia ntau yam proteins nyob rau tib lub sijhawm siv txoj kev tsis muaj cell (piv txwv li, kuaj cov protein hloov pauv los ntawm cov lus me me los ntawm ntau cov qauv DNA sib txawv). Hauv qhov kev sim no tus neeg sawv cev, IVT system tau siv los qhia txog tib neeg cov protein caspase-3.
Cov lus xaus thiab lub zeem muag yav tom ntej
Kev tsim cov protein ntau tam sim no tuaj yeem pom tias yog kev qhuab qhia paub tab. Qhov no yog qhov tshwm sim ntawm ntau qhov kev txhim kho ntxiv hauv kev ua kom huv thiab tshuaj xyuas. Tam sim no, cov kev pab cuam tshawb nrhiav tshuaj tsis tshua raug tso tseg vim tsis muaj peev xwm tsim cov hom phiaj protein. Cov txheej txheem sib npaug rau kev qhia, kev ua kom huv thiab kev tshuaj xyuas ntawm ntau cov tshuaj recombinant tam sim no paub zoo nyob rau hauv ntau lub chaw soj nstuam thoob ntiaj teb.
Protein complexes thiab loj hlob kev vam meej hauv kev uasolubilized membrane cov qauv yuav xav tau ntau qhov kev hloov pauv kom raws li qhov xav tau. Kev tshwm sim ntawm cov koom haum tshawb fawb tau txais txiaj ntsig rau kev muab cov protein ntau ntxiv yuav tso cai rau kev muab cov peev txheej tshawb fawb kom tau raws li cov kev cov nyom tshiab no.
Tsis tas li ntawd, kev ua haujlwm sib npaug yuav tsum tso cai tsim cov tsev qiv ntawv ua tiav ntawm cov tshuaj saib xyuas kom muaj kev txheeb xyuas lub hom phiaj tshiab thiab kev tshuaj ntsuam xyuas qib siab, nrog rau cov tshuaj me me ntawm cov tshuaj tshawb pom.
