Qhov sib txawv tseem ceeb ntawm GFP thiab EGFP yog tias GFP yog hom tsiaj qus uas muaj nyob rau hauv cov molecular cloning ntawm cov hlwb uas tsis yog tsiaj txhu thaum lub EGFP yog ib qho kev txhim kho lossis kev tsim qauv ntawm GFP uas tuaj yeem siv rau ntawm cov tsiaj txhu..
Molecular cloning yog cov txheej txheem siab heev uas cov kws tshawb fawb siv ntau heev hauv kev nthuav qhia cov proteins los ntawm kev siv thev naus laus zis. Hauv recombinant DNA thev naus laus zis, nws yog ib qho tsim nyog los hloov cov vector rov ua kom tiav los tuav lub cev. Yog li ntawd, thaum lub sij hawm kev hloov pauv, nws yuav tsum raug txheeb xyuas thiab paub tseeb tias cov noob ntawm kev txaus siab tau hloov pauv lossis tsis yog rau tus tswv tsev. Txhawm rau ntsuas qhov no, molecular biologists tau txais ntau txoj kev. Tawm ntawm cov txheej txheem, ib qho yog reporter gene. Cov noob neeg sau xov xwm no ua raws li cov cim xaiv tau los xaiv qhov tseeb hloov pauv. Yog li, Green Fluorescent Protein (GFP) thiab Enhanced Green Fluorescent Protein (EGFP) yog ob tus neeg tshaj tawm cov proteins uas siv rau hauv molecular cloning.
GFP yog dab tsi?
GFP yog hom tsiaj qus uas muaj 238 amino acid residues thiab ob peb cheeb tsam xaiv tau ntawm cov amino acid sequences uas txawv ntawm lwm cov fluorescent proteins. Tsis tas li ntawd, hom tsiaj qus no yog thawj zaug nyob rau hauv kev sib cais los ntawm Aequorea Victoria; ib hom jellyfish. Txawm li cas los xij, hauv cov xwm txheej ntuj, cov jellyfish muaj peev xwm tsim cov xim ntsuab fluorescence los teb rau qee yam stimuli.
Thaum ntxov, lub tswv yim no ua rau cov kws tshawb fawb xav tsis thoob, thiab lawv tau txiav txim siab siv nws ntawm lawv cov thev naus laus zis DNA sib txuas. Yog li ntawd, cov kws tshawb fawb tau siv daim ntawv hloov pauv ntawm cov noob tsiaj qus los ua cov neeg sau xov xwm gene hauv lawv cov kev tshawb fawb gene. Hom tsiaj qus ntawm GFP muaj peev xwm tsim tau cov protein uas muab fluorescence ntawm chav tsev kub lossis hauv qab UV teeb. Yog li ntawd, thaum muab tso rau hauv lub transformants, nws qhia thiab tsim fluorescence. Yog tias cov fluorescence tshwm sim tom qab cov txheej txheem hloov pauv, nws lees paub qhov ua tiav ntawm txoj kev hloov pauv. Hauv cov lus yooj yim, fluorescence emission qhia qhov kev hloov pauv ntawm vector uas nqa cov noob paj rau hauv tus tswv.
Daim duab 01: GFP
Vim yog vim li no, GFP ua raws li tus cim hauv vivo ntawm cov noob qhia. Tam sim no, genetic engineering cov txheej txheem yog siv los tsim GFP. Tsis tas li ntawd, muaj ntau yam kev txhim kho ntawm GFP xws li EGFP. Li no, qhov no ua rau muaj txiaj ntsig zoo ntawm GFP hauv kev tshawb fawb molecular cloning thiab gene qhia.
EGFP yog dab tsi?
Txhim kho ntsuab Fluorescent Protein lossis EGFP yog ib qho kev txhim kho ntawm GFP. Hauv cov lus yooj yooj yim, peb tuaj yeem txhais EGFP raws li kev tsim qauv ntawm cov tsiaj qus GFP. Thaum cov noob tsiaj qus ntawm GFP hloov pauv, nws ua rau muaj txiaj ntsig zoo. Li no, cov noob hloov pauv ntawm GFP tso cai rau kev nthuav tawm cov cim tshiab, thiab vim li ntawd, peb tuaj yeem tsim kho GFP nrog cov yam ntxwv zoo dua. Tsis tas li ntawd, peb tuaj yeem qhia txog kev hloov pauv tau zoo rau cov tsiaj qus-hom GFP gene siv irradiation lossis chemical method. Cov noob hloov pauv no ces tsim cov EGFP, uas muaj cov yam ntxwv zoo dua.
Figure 02: EGFP
Cov yam ntxwv zoo dua ntawm EGFP yog raws li hauv qab no;
- Muaj peev xwm tshaj tawm cov teeb liab muaj zog fluorescence.
- muaj siab rhiab heev.
- Tau siv tau rau ntawm cov kabmob mammalian es tsis txhob prokaryotes thiab lwm yam eukaryotes qis.
- Kuj, muab kev ua kom huv huv ntawm cov khoom.
Yog li ntawd, piv rau GFP, EGFP yog qhov kev xaiv zoo tshaj plaws rau kev tshawb fawb txog noob caj noob ces. Txawm li cas los xij, cov khoom lag luam kim dua piv rau GFP.
Dab tsi yog qhov zoo sib xws ntawm GFP thiab EGFP?
- GFP thiab EGFP yog ob lub proteins uas muaj peev xwm ua kom muaj xim ntsuab
- Yog li ntawd, ob qho tib si ua haujlwm raws li tus neeg sau xov xwm cov proteins hauv cov kev tshawb fawb gene qhia.
- Kuj, nws tuaj yeem ua rau ob qho tib si siv cov thev naus laus zis DNA recombinant.
- Ntxiv mus, nws yog qhov yooj yim los hloov pauv ob hom no los ua ke cov ntaub ntawv txhim kho.
Qhov txawv ntawm GFP thiab EGFP yog dab tsi?
Tus neeg sau xov xwm gene yog ib lub noob uas txuas nrog cov noob ntawm kev txaus siab nyob rau hauv recombinant DNA technology. Nws qhia txog kev ua tiav kev hloov pauv ntawm cov vector recombinant rau tus tswv tsev. Ntawm no, GFP thiab EGFP yog ob hom ntsuab florescent proteins uas ua hauj lwm raws li reporter proteins. Txawm li cas los xij, qhov sib txawv tseem ceeb ntawm GFP thiab EGFP yog tias GFP yog hom tsiaj qus thaum EGFP yog ib qho qauv tsim ntawm GFP. Tsis tas li ntawd, EGFP muaj cov yam ntxwv zoo dua li GFP. Piv txwv li, EGFP tsim lub teeb muaj zog fluorescent thiab muaj kev nkag siab ntau dua li GFP. Lwm qhov sib txawv ntawm GFP thiab EGFP yog cov tshuab uas peb tuaj yeem siv tau. Cov kab mob uas tsis yog tsiaj txhu siv GFP thaum cov tsiaj txhu siv EGFP.
Cov duab hauv qab no nthuav tawm qhov sib txawv ntawm GFP thiab EGFP hauv daim ntawv teev npe.
Summary – GFP vs EGFP
GFP thiab EGFP yog cov neeg sau xov xwm cov proteins hauv molecular cloning thiab gene qhia kev tshawb fawb. GFP yog hom tsiaj qus, uas yog ntsuab fluorescent protein. Cov protein tau pib nyob rau hauv kev sib cais los ntawm jellyfish Aequorea victoria. Hauv qhov sib piv, EFGP yog ib qho kev txhim kho ntawm GFP protein. Nws yog ib tug mutant ntawm cov qus-hom nrog txhim kho cov yam ntxwv. Li no, EFGP muaj lub teeb liab siab zog thiab siab dua. Yog li ntawd, peb tuaj yeem siv nws ntawm cov tsiaj txhu tsiaj. Hauv qhov sib piv, kev siv GFP feem ntau tsuas yog ntawm cov tsiaj tsis yog tsiaj txhu. Nyob rau hauv tag nrho, qhov no yog qhov txawv ntawm GFP thiab EGFP.
