Qhov sib txawv tseem ceeb ntawm polysome profileing thiab ribosome profiling yog tias polysome profiling txheeb xyuas tus cwj pwm ribosome siv ob qho tib si ribosome thiab mRNA (polysome) thaum lub sij hawm txhais lus, thaum ribosome profiling txheeb xyuas tus cwj pwm ribosome tsuas yog siv mRNA ib ntus thaum txhais lus.
Kev txhais lus yog theem thib ob ntawm kev sib txuas ntawm cov protein uas hloov cov ntaub ntawv hauv mRNA mus rau ib qho amino acid sequence. Translatomics yog ib qho kev kawm ntawm ORFs (qhib cov ntawv nyeem) uas nquag txhais hauv ib lub xov tooj ntawm lub cev. Polysome thiab ribosome profiling cov tswv yim yog ob hom kev kawm hauv thaj tsam ntawm molecular biology los ntsuas thiab ntsuas qhov sib txawv ntawm cov ntsiab lus ntawm kev tshuaj xyuas cov txhais lus.
Polysome Profileing yog dab tsi?
Polysome profileing yog ib qho txheej txheem uas cuam tshuam cov xwm txheej ntawm kev txhais lus ntawm mRNA tshwj xeeb los ntawm kev tshuaj xyuas tus cwj pwm ntawm ob qho tib si ribosome thiab mRNA (polysome). Hauv lwm lo lus, cov txheej txheem no muab cov ntaub ntawv thiab cov lus xaus ntawm kev koom tes ntawm mRNAs nrog ribosomes. Lub polysome hais txog cov pab pawg ntawm ribosomes khi rau ib qho mRNA uas muaj nyob rau lub sijhawm elongation theem ntawm kev txhais lus.
Daim duab 01: Polysome Profileing
Polysome profileing xav tau lub cell lysate, uas yog ces centrifuged. Cov qauv centrifuged yog tom qab ntawd sib cais raws li lawv qhov ntom ntom kom paub qhov me me thiab loj subunits ntawm ribosomes thiab sib thooj mRNA koom nrog hauv kev tsim cov polysome. Tsis tas li ntawd, cov txheej txheem kuj suav nrog ntsuas qhov muag pom qhov ntom ntom. Cov kws tshaj lij yuav tsum tau ua kom muaj kev sib txuas ntawm polysome.
Polysome profileing txheej txheem yog ib qho cuab yeej tseem ceeb rau ntau daim ntawv thov. Cov kws tshawb fawb siv cov txheej txheem no los kawm txog qib kev txhais lus hauv hlwb. Tshwj xeeb tshaj yog, nws yog ib qho cuab yeej los muab cov ntaub ntawv tseeb ntawm kev kawm ntawm tus kheej cov proteins thiab lawv cov mRNAs tshwj xeeb. Hauv cov ntsiab lus ntawm kev kawm qib kev txhais lus ntawm ib qho mRNA, cov txheej txheem polysome profile yog qhov tseem ceeb. Ntawm no, qhov 3' thiab 5' ib ntus ntawm mRNA tuaj yeem tshawb xyuas nrog kev siv rau lawv cov teebmeem ntawm tus nqi ntawm mRNA tsim thiab qib kev txhais lus.
Ribosome Profiling yog dab tsi?
Ribosome profiling yog ib qho txheej txheem uas txheeb xyuas tus cwj pwm ntawm ribosome nrog rau nws mRNA thaum txhais lus. Cov txheej txheem no tau pom thiab tsim los ntawm Joan Steitz thiab Marilyn Kozak. Tom qab no cov cuab yeej no tau tsim ntxiv los ntawm ob tus kws tshawb fawb, Nicholas thiab Jonathan, ua ke nrog cov txheej txheem tom ntej, uas ua rau muaj kev txhim kho ntawm cov txheej txheem sib txawv xws li Kev Txhais Ribosome Affinity Purification (TRAP) txoj kev.
Daim duab 02: Ribosome Sequencing
Cov txheej txheem ntawm ribosome profileing suav nrog kev cais mRNA, tshem RNA uas tsis raug khi rau ribosomes, thiab cais mRNA khi rau ribosomes. Ua raws li cov txheej txheem no, mRNA cais tawm yog thim rov qab, thiab cDNA synthesis tshwm sim. Thaum kawg, cov ntaub ntawv ua ntu zus tuaj yeem ua raws li cov lus qhia kev txhais lus los txiav txim siab tus yam ntxwv ntawm tus cwj pwm ribosome nrog rau mRNA.
Ribosome profiling pab ntau tus kws tshawb fawb los txheeb xyuas thiab txheeb xyuas qhov chaw pib ntawm kev txhais lus, kev ua tiav ntawm cov lus qhib kev nyeem ntawv (ORFs) hauv lub xov tooj ntawm tes lossis cov ntaub so ntswg, kev faib tawm ntawm ribosomes ntawm mRNA, thiab tus nqi ntawm txhais ribosomes. Ribosome profiling tseem hu ua ribosome footprinting lossis Ribo Seq.
Dab tsi yog qhov zoo sib xws ntawm Polysome Profiling thiab Ribosome Profiling?
- Polysome thiab ribosome profileing yog cov txheej txheem lom neeg molecular tseem ceeb hauv kev tshawb fawb.
- Lawv muab lus qhia txog txheej txheem txhais lus.
- Ob qhov txheej txheem profile muab cov ntaub ntawv los ntawm kev tshuaj xyuas ntawm kev txhais lus.
- yuav tsum muaj cov kws tshaj lij los ua ob txoj hauv kev kom tau txais qhov tseeb.
- cov cuab yeej Bioinformatics ua lub luag haujlwm tseem ceeb hauv ob txoj kev.
Qhov txawv ntawm Polysome Profiling thiab Ribosome Profiling yog dab tsi?
Polysome profiling txheeb xyuas tus cwj pwm ribosome siv ob qho tib si ribosome thiab mRNA (polysome) thaum lub sij hawm txhais lus, thaum ribosome profiling txheeb xyuas tus cwj pwm ribosome tsuas yog siv mRNA ib ntus thaum txhais lus. Yog li, qhov no yog qhov sib txawv tseem ceeb ntawm polysome profileing thiab ribosome profileing. Tsis tas li ntawd, polysome profileing suav nrog cov txheej txheem xws li qhov ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom siv mRNA cov txheej txheem sib sib zog ua ke. Tsis tas li ntawd, ribosome profileing yog qhov tseeb dua li polysome profileing.
Cov hauv qab no infographic nthuav tawm qhov sib txawv ntawm polysome thiab ribosome profileing hauv daim ntawv tabular rau ib sab ntawm kev sib piv.
Summary – Polysome Profiling vs Ribosome Profiling
Kev txhais lus yog theem thib ob ntawm kev tsim cov protein uas cuam tshuam nrog kev hloov cov ntaub ntawv ntawm mRNA ib ntus mus rau ib qho amino acid sequence. Cov txheej txheem no yuav tsum muaj mRNA template, ribosomes, amino acids, tRNA, thiab lwm yam. Polysome thiab ribosome profileing yog ob txoj hauv kev molecular. Polysome profiling txheeb xyuas tus cwj pwm ribosome siv ob qho tib si ribosome thiab mRNA (polysome) thaum lub sij hawm txhais lus, thaum ribosome profiling txheeb xyuas tus cwj pwm ribosome tsuas yog siv mRNA sib lawv liag thaum txhais lus. Polysome profileing muaj cov txheej txheem xws li qhov ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom ntom. Ribosome profileing muaj cov txheej txheem xws li mRNA extraction, cDNA synthesis, thiab sequencing. Yog li, qhov no qhia txog qhov sib txawv ntawm polysome profileing thiab ribosome profileing.
