Qhov sib txawv tseem ceeb ntawm cov poov xab ib hybrid thiab poov xab ob hybrid yog cov poov xab ib hybrid yog ib qho hauv vitro raws li kev ntsuam xyuas los soj ntsuam DNA-protein kev sib cuam tshuam thaum poov xab ob hybrid yog ib qho kev ntsuam xyuas hauv vitro los soj ntsuam cov protein-protein kev sib cuam tshuam.
Kawm txog kev sib cuam tshuam ntawm cov cellular molecules tuaj yeem pab txhais cov chaw paub txog kev koom tes hauv cov noob caj noob ces thiab txoj hauv kev protein. Kev sib cuam tshuam ntawm DNA thiab protein yog ib qho tseem ceeb rau cov txheej txheem ntawm tes xws li DNA hloov kho thiab kev hloov pauv hloov pauv, thiab lwm yam. Ntawm qhov tod tes, kev sib cuam tshuam ntawm cov protein thiab cov protein ua lub luag haujlwm tseem ceeb hauv cov txheej txheem ntawm tes xws li kev rov ua dua, kev hloov pauv, kev txhais lus, thiab cov teeb liab hloov. Yog li ntawd, nws yog ib qho tseem ceeb heev uas yuav tau txheeb xyuas cov kev sib cuam tshuam ntawm cellular molecules los ntawm kev ntsuam xyuas sib txawv. Poov ib hybrid thiab poov xab ob hybrid yog ob nyob rau hauv vitro raws li kev soj ntsuam los soj ntsuam DNA-protein thiab protein-protein sib cuam tshuam.
Yeast Ib Hybrid yog dab tsi?
Yeast ib hybrid (Y1H) yog ib qho kev soj ntsuam hauv vitro los soj ntsuam kev sib cuam tshuam hauv cov cellular DNA-protein. Txoj kev no yog thawj zaug tsim los ntawm Wang thiab Reed hauv xyoo 1993. Txij thaum ntawd los, nws tau pom lub zog loj dua hauv kev tshawb fawb txog roj ntsha. Hauv qhov kev ntsuam xyuas no, muaj ob lub ntsiab lus tseem ceeb: DNA bait thiab protein preys.
Daim duab 01: Yeast Ib Hybrid
Nyob rau hauv cov txheej txheem, DNA ib ntus (DNA bait) yog thawj cloned upstream ntawm ob tus neeg sau xov xwm sib txawv (HIS3, LacZ). Cov neeg sau xov xwm no tau tsim ua ke rau hauv genome ntawm cov kab mob poov xab. Cov kev hloov pauv tau raug coj los ua cov protein preys. Cov txheej txheem kev hloov pauv yog fused mus rau transcriptional activation domain (AD) ntawm cov poov xab Gal4 transcriptional kom thiaj li ua tau cov protein ntau. Tag nrho cov hybrid protein no tau muab tso rau hauv plasmid ua ntej qhia nws mus rau hauv cov kab mob poov xab. Tom qab qhov kev taw qhia ntawm cov plasmid recombinant, thaum lub transcriptional yam cuam tshuam nrog DNA fragment, AD moiety activates tus reporter noob qhia (HIS3or LacZ). Sequencing lub plasmids tso cai rau kev txheeb xyuas ntawm qhov transcriptional yam uas tuaj yeem khi rau DNA tawg. Tsis ntev los no, poov xab ib txoj kev hybrid tau siv los tshawb nrhiav kab mob. Yog li ntawd, yav tom ntej, qhov kev ntsuam xyuas no yuav siv dav dua hauv kev tshawb fawb thiab kev kho mob.
Yeast Ob Hybrid yog dab tsi?
Yeast ob hybrid (Y2H) yog ib qho kev ntsuam xyuas hauv vitro los txheeb xyuas cov protein thiab cov protein sib cuam tshuam. Txoj kev no yog thawj zaug pom los ntawm Stanley Fields thiab Ok-Kyu Song hauv xyoo 1989. Cov txheej txheem no kuaj pom cov protein-protein sib cuam tshuam siv Gal4 transcription yam ntawm cov poov xab.
Daim duab 02: Poov ob hybrid
Nyob rau hauv txoj kev no, ob lub fusions (hybrids) tau tsim ua ntej siv cov proteins uas txaus siab: fusing ib cov protein nrog DNA binding domain (DBD) thiab los ntawm kev sib txuas nrog lwm cov protein nrog cov activator domain (AD) ntawm Gal4 transcription factor. Cov protein fused nrog DBD yog hu ua bait thaum, cov protein fused nrog AD yog hu ua prey. Hauv cov kab mob poov xab, raws li kev sib cuam tshuam ntawm cov kab nuv ntses thiab cov tsiaj nyeg, DBD thiab AD raug coj los ze ze los tsim kom muaj kev ua haujlwm ntawm Gal4 transcription tshaj tawm ntawm cov neeg sau xov xwm gene. Lub functional transcription factor activates reporter gene qhia. Tsis tas li ntawd, txoj kev no yog tam sim no siv los tshuaj xyuas cov kev sib cuam tshuam ntawm ib qho membrane proteins nyob rau hauv mammalian hlwb.
Yeast Ib hybrid thiab Poov ob hybrid yog dab tsi?
- Poj niam ib tug hybrid thiab poov xab ob hybrid yog ob nyob rau hauv vitro raws li kev soj ntsuam los soj ntsuam DNA-protein thiab protein-protein sib cuam tshuam.
- Cov kev ntsuas no siv cov kab mob poov xab.
- Ob qhov kev ntsuam xyuas siv qhov ua kom ua haujlwm (AD) ntawm Gal4 qhov kev hloov pauv.
- Lawv siv cov neeg sau xov xwm gene (HIS3, lossis LacZ) qhia kom pom kev sib cuam tshuam.
- Lawv muaj baum thiab prey.
- Ob qhov kev ntsuam xyuas no muaj txiaj ntsig zoo hauv kev tshawb fawb lom neeg niaj hnub no.
Yeast Ib hybrid thiab Poov ob hybrid yog dab tsi?
Yeast ib hybrid yog ib qho kev ntsuam xyuas hauv vitro los soj ntsuam DNA thiab cov protein sib cuam tshuam thaum cov poov xab ob hybrid yog ib qho kev ntsuam xyuas hauv vitro los txheeb xyuas cov protein thiab cov protein sib cuam tshuam. Yog li, qhov no yog qhov sib txawv ntawm cov poov xab ib hybrid thiab poov xab ob hybrid. Tsis tas li ntawd, nyob rau hauv poov xab ib hybrid, bait yog ib tug DNA tawg, thiab prey yog ib tug protein molecule. Ntawm qhov tod tes, nyob rau hauv cov poov xab ob hybrid, ob qho tib si bait thiab prey yog protein molecules.
Cov hauv qab no infographic teev qhov sib txawv ntawm cov poov xab ib hybrid thiab poov xab ob hybrid nyob rau hauv daim ntawv tabular rau ib sab sib piv.
Summary – Poov ib hybrid vs poov ob hybrid
Poj niam ib hybrid thiab poov xab ob hybrid yog ob qho tseem ceeb hauv vitro raws li kev soj ntsuam. Poov ib tug hybrid yog ib qho kev ntsuam xyuas hauv vitro los txheeb xyuas DNA thiab cov protein sib cuam tshuam thaum cov poov xab ob hybrid yog ib qho kev ntsuam xyuas hauv vitro los soj ntsuam cov protein thiab cov protein sib cuam tshuam. Yog li, qhov no yog qhov sib txawv ntawm cov poov xab ib hybrid thiab poov xab ob hybrid.
